NUCLEIC ACID MOLECULES AND OTHER MOLECULES ASSOCIATED WITH THE TETRAPYRROLE PATHWAY

Abstract

in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, nucleic acid sequences from maize and soybean associated with the tetrapyrrole pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001]This application is a continuation of U.S. application Ser. No. 09/233,218 filed Jan. 20, 1999, which is a continuation-in-part of U.S. application Ser. No. 09/198,779, filed Nov. 24, 1998 (abandoned), which claims the benefit of U.S. Provisional Appln. Ser. No. 60/067,000, filed Nov. 24, 1997; and to U.S. Provisional Appln. Ser. No. 60/066,873, filed Nov. 25, 1997; and to U.S. Provisional Appln. Ser. No. 60/069,472, filed Dec. 9, 1997; and to U.S. Provisional Appln. Ser. No. 60/074,201, filed Feb. 10, 1998; and to U.S. Provisional Appln. Ser. No. 60/074,280, filed Feb. 10, 1998; and to U.S. Provisional Appln. Ser. No. 60/074,281, filed Feb. 10, 1998; and to U.S. Provisional Appln. Ser. No. 60/074,282, filed Feb. 10, 1998; and to U.S. Provisional Appln. Ser. No. 60/074,565, filed Feb. 12, 1998; and to U.S. Provisional Appln. Ser. No. 60/074,566, filed Feb. 12, 1998; and to U.S. Provisional Appln. Ser. No. 60/074,567, filed Feb. 12, 1998; and to U.S. Provisional Appln. Ser. No. 60/074,789, filed Feb. 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/075,459, filed Feb. 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/075,460, filed Feb. 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/075,461, filed Feb. 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/075,462, filed Feb. 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/075,463, filed Feb. 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/075,464, filed Feb. 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/077,229, filed Mar. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/077,230, filed Mar. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/077,231, filed Mar. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/078,031, filed Mar. 16, 1998; and to U.S. Provisional Appln. Ser. No. 60/078,368, filed Mar. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/080,844, filed Apr. 7, 1998; and to U.S. Provisional Appln. Ser. No. 60/083,067, filed Apr. 27, 1998; and to U.S. Provisional Appln. Ser. No. 60/083,386, filed Apr. 29, 1998; and to U.S. Provisional Appln. Ser. No. 60/083,387, filed Apr. 29, 1998; and to U.S. Provisional Appln. Ser. No. 60/083,388, filed Apr. 29, 1998; and to U.S. Provisional Appln. Ser. No. 60/083,389, filed Apr. 29, 1998; and to U.S. Provisional Appln. Ser. No. 60/084,684, filed May 8, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,222, filed May 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,223, filed May 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,224, filed May 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,245, filed May 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,183, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,184, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,185, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,186, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,187, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,188, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,339, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,524, filed Jun. 16, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,810, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,814, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/091,247, filed Jun. 30, 1998; and to U.S. Provisional Appln. Ser. No. 60/092,036, filed Jul. 8, 1998; and to U.S. Provisional Appln. Ser. No. 60/099,667, filed Sep. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/099,668, filed Sep. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/099,670, filed Sep. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/099,697, filed Sep. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/100,672, filed Sep. 16, 1998; and to U.S. Provisional Appln. Ser. No. 60/100,673, filed Sep. 16, 1998; and to U.S. Provisional Appln. Ser. No. 60/100,674, filed Sep. 16, 1998; and to U.S. Provisional Appln. Ser. No. 60/101,130, filed Sep. 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/101,132, filed Sep. 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/108,996, filed Nov. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/109,018, filed Nov. 18, 1998. Application Ser. No. 09/233,218 is also a continuation-in-part of U.S. application Ser. No. 09/227,586, filed Jan. 8, 1999 (abandoned), which claims the benefit of U.S. Provisional Appln. Ser. No. 60/071,064, filed Jan. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/090,170, filed Jun. 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/092,036, filed Jul. 8, 1998. Application Ser. No. 09/233,218 is also a continuation-in-part of U.S. application Ser. No. 09/229,413, filed Jan. 12, 1999 (abandoned), which claims the benefit of U.S. Appln. Ser. No. 60/071,479, filed Jan. 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,533, filed May 15, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,806, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,807, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,808, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,811, filed Jun. 18, 1998; and to U.S.

Provisional Appln. Ser. No. 60/089,812, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,813, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/091,247, filed Jun. 30, 1998; and to U.S. Provisional Appln. Ser. No. 60/091,405, filed Jun. 30, 1998; and to U.S. Provisional Appln. Ser. No. 60/099,697, filed Sep. 9, 1998; and to U.S. Provisional Appln. Ser. No. 60/100,963, filed Sep. 17, 1998; and to U.S. Provisional Appln. Ser. No. 60/101,343, filed Sep. 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/101,344, filed Sep. 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/101,347, filed Sep. 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/101,508, filed Sep. 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/101,707, filed Sep. 25, 1998; and to U.S. Provisional Appln. Ser. No. 60/104,124, filed Oct. 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/104,126, filed Oct. 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/104,127, filed Oct. 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/104,128, filed Oct. 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/111,981, filed Dec. 11, 1998. Application Ser. No. 09/233,218 also claims the benefit of U.S. Provisional Appln. Ser. No. 60/072,027, filed Jan. 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/072,888, filed Jan. 27, 1998; and to U.S. Provisional Appln. Ser. No. 60/076,709, filed Mar. 4, 1998; and to U.S. Provisional Appln. Ser. No. 60/076,912, filed Mar. 6, 1998; and to U.S. Provisional Appln. Ser. No. 60/078,031, filed Mar. 16, 1998; and to U.S. Provisional Appln. Ser. No. 60/083,390, filed Apr. 29, 1998; and to U.S. Provisional Appln. Ser. No. 60/084,684, filed May 8, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,057, filed May 8, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,429, filed May 8, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,245, filed May 13, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,533, filed May 15, 1998; and to U.S. Provisional Appln. Ser. No. 60/085,940, filed May 19, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,339, filed May 21, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,594, filed May 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/086,608, filed May 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/087,422, filed Jun. 1, 1998; and to U.S. Provisional Appln. Ser. No. 60/087,631, filed Jun. 2, 1998; and to U.S. Provisional Appln. Ser. No. 60/087,762, filed Jun. 2, 1998; and to U.S. Provisional Appln. Ser. No. 60/087,972 filed Jun. 4, 1998; and to U.S. Provisional Appln. Ser. No. 60/087,973 filed Jun. 4, 1998; and to U.S. Provisional Appln. Ser. No. 60/088,441, filed Jun. 8, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,627, filed Jun. 16, 1998; and to U.S. Provisional Appln. Ser. No. 60/089,789, filed Jun. 18, 1998; and to U.S. Provisional Appln. Ser. No. 60/090,170, filed Jun. 22, 1998; and to U.S. Provisional Appln. Ser. No. 60/090,856, filed Jun. 26, 1998; and to U.S. Provisional Appln. Ser. No. 60/090,928, filed Jun. 26, 1998; and to U.S. Provisional Appln. Ser. No. 60/091,035, filed Jun. 29, 1998; and to U.S. Provisional Appln. Ser. No. 60/091,247, filed Jun. 30, 1998; and to U.S. Provisional Appln. Ser. No. 60/091,405, filed Jun. 30, 1998; and to U.S. Provisional Appln. Ser. No. 60/092,036, filed Jul. 8, 1998; and to U.S. Provisional Appln. Ser. No. 60/100,963, filed Sep. 17, 1998; and to U.S. Provisional Appln. Ser. No. 60/110,108, filed Nov. 25, 1998; and to U.S. Provisional Appln. Ser. No. 60/110,109, filed Nov. 25, 1998; and to U.S. Provisional Appln. Ser. No. 60/111,033, filed Dec. 4, 1998; and to U.S. Provisional Appln. Ser. No. 60/111,742, filed Dec. 10, 1998. All of the above-listed applications are herein incorporated by reference in their entirety.

INCORPORATION OF SEQUENCE LISTING

[0002]This application contains an electronic equivalent paper copy of the sequence listing submitted herewith electronically via EFS web and a computer-readable form of the sequence listing submitted herewith electronically via EFS web and contains the file named "P30114US04_seqlist.txt", which is 49,918,550 bytes in size (measured in MS-DOS) and which was also created on Jun. 11, 2009.

FIELD OF THE INVENTION

[0003]The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, nucleic acid sequences from maize and soybean plants associated with the tetrapyrrole pathway in plants. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

BACKGROUND OF THE INVENTION

[0004]I. Biosynthesis of Tetrapyrroles

[0005]The biosynthesis of tetrapyrroles such as heme and chlorophyll as well as a number of other tetrapyrroles such as siroheme, the cofactor for sulfite and nitrite reductases, cobalamin (vitamin B12), and the chromophore of phytochrome, can be subdivided into three major phases; ALA synthesis, porphyrin ring synthesis and synthesis of final products. The pathway is conserved among species except for the synthesis of 5-aminolevulinate, also known as 5-aminolevulinic acid ("ALA") (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995), both of which are herein incorporated by reference).

[0006]The first phase of the biosynthesis of tetrapyrroles, such as heme and chlorophyll, is the synthesis of ALA. Yeast, fungi, mammals and some bacteria (the α-group of proteobacteria or purple bacteria, e.g. Bradyrhizobium japonicum and Rhodobacter capsulatus) biosynthesize tetrapyrroles via the single step four-carbon (C4), or Shemin pathway. In this pathway ALA synthase (E.C. 2.3.1.37) catalyzes the condensation of glycine with succinyl-CoA to generate ALA.

[0007]Plants, green algae, cyanobacteria, most eubacteria (e.g. E. coli and Bacillus subtilis), and archaebacteria biosynthesize ALA via the three-step five-carbon ("C5") pathway, which includes glutamyl-tRNA synthetase ("GluRS"), glutamyl-tRNA reductase ("GluTR") and glutamate-1-semialdehyde aminotransferase ("GSA-AT"). In plants and algae, the C5 pathway is localized in the chloroplast. The formation of ALA via the C5 pathway is reported to be the rate-limiting step in the biosynthesis of heme and chlorophyll (Kumar et al., Trends in Plant Science 1:371-376 (1996); Tanaka et al., Plant Physiol. 110:1223-30 (1996); Masuda et al., Plant Physiol. Biochem. 34:11-16 (1996); Hungerer et al., J. Bacteriol. 177:1435-43 (1995); Ilag et al., Plant Cell 6:265-75 (1994), all of which are herein incorporated by reference in their entirety).

[0008]Chloroplastic GluRS (E.C. 6.1.1.17), also known as glutamate-tRNA ligase, converts glutamate to glutamyl-tRNA ("Glu-tRNA") activating the C-1 of glutamate in an ATP dependent reaction (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). Glu-tRNA is reported to be the first intermediate in the C5 pathway and it also reported to serve as a source of glutamate in protein biosynthesis. GluRS is a soluble plastid enzyme which has been isolated from higher plants (barley, wheat) and other organisms. Reported GluRS enzymes are homodimers encoded by a nuclear gene and synthesized in the cytoplasm and have a molecular weight of 54 kD (barley) and 56 kD (wheat).

[0009]GluTR, the first committed enzyme reported in heme and chlorophyll biosynthesis, catalyzes the NADPH dependent reduction of Glu-tRNA to glutamate 1-semialdehyde ("GSA") with the release of intact tRNA (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). GluTR is reported as the rate limiting step in ALA formation and is present only at low levels in all organisms examined (Masuda et al., Plant Physiol. Biochem. 34:11-16 (1996); Schroeder et al., Biochem. J. 281:843-50 (1992), the entirety of which is herein incorporated by reference; Masuda et al., Plant Cell Physiol. 36:1237-43 (1995), the entirety of which is herein incorporated by reference). Plant GluTR is a soluble enzyme localized in plastids and encoded in the nucleus. GluTR has been reported to exist as a multimer of a single subunit. The purified barley enzyme has a molecular weight of 270 kD with a monomeric subunit size of 54 kD (Pontoppidan and Kannangara, Eur. J. Biochem. 225:529-37 (1994), the entirety of which is herein incorporated by reference). Arabidopsis and cucumber enzymes have similar subunit molecular weights (Tanaka et al., Plant Physiol. 110:1223-30 (1996); Ilag et al., Plant Cell 6:265-75 (1994); Kumar et al., Plant Mol. Biol. 30:419-26 (1996), the entirety of which is herein incorporated by reference).

[0010]GluTR genes (also known as HEMA genes) have been cloned and the amino acid sequences determined for a number of sources including three higher plants; Arabidopsis, barley, and cucumber. The deduced amino acid sequence of GluTR from all sources exhibit about 60% overall similarity with stretches of amino acid identity. Barley, Arabidopsis, and cucumber show over 70% identity at the deduced amino acid level (Vothknecht et al., Proc. Natl. Acad. Sci. (U.S.A.) 93:9287-9291 (1996), the entirety of which is herein incorporated by reference). Two different GluTR genes have been isolated from three higher plants; Arabidopsis (Ilag et al., Plant Cell 6:265-75 (1994)), barley (Bougri and Grimm, Plant J. 9:867-878 (1996), the entirety of which is herein incorporated by reference), and cucumber (Masuda et al., Plant Cell Physiol. 36:1237-43 (1995), the entirety of which is herein incorporated by reference). In Arabidopsis and cucumber, one GluTR gene is expressed in all tissues and a second is expressed in a tissue specific manner. These genes are also reported to be differentially regulated by light (Tanaka et al., Plant Physiol. 110:1223-30 (1996); Masuda et al., Plant Physiol. Biochem. 34:11-16 (1996); Ilag et al., Plant Cell 6:265-75 (1994); Masuda et al., Plant Cell Physiol. 36:1237-43 (1995); Kumar et al., Plant Mol. Biol. 30:419-26 (1996); Hori et al., Plant Physiol. Biochem. 34:3-9 (1996), the entirety of which is herein incorporated by reference).

[0011]GSA-AT (glutamate-1-semialdehyde aminotransferase (E.C. 5.4.3.8)), catalyzes the conversion of GSA to ALA. GSA-AT is a soluble protein localized in the chloroplast and encoded in the nucleus (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). It has a subunit molecular weight of about 45 kD. The holoenzyme consists of two identical subunits and utilizes pyridoxal phosphate ("PLP") as a cofactor (Kumar et al., Trends in Plant Science 1:371-376 (1996); Gough et al., Glutamate 1-semialdehyde aminotransferase as a target for herbicides, Boeger, Ed., Lewis, Boca Raton, Fla., (1993), the entirety of which is herein incorporated by reference). GSA-AT is reported to be inhibited by gabaculine, which has also been shown to inhibit chlorophyll biosynthesis in barley leaves (Rogers and Smith, BCPC Monogr. 42:183-93 (1989), the entirety of which is herein incorporated by reference). GSA-AT has been crystallized from Synechococcus (Hennig et al., J. Mol. Biol. 242:591-594 (1994); Hennig et al., Proc. Natl. Acad. Sci. (U.S.A.) 94:4866-4871 (1997), both of which are herein incorporated by reference in their entirety).

[0012]GSA-AT genes have been cloned from a number of plants including Arabidopsis. The deduced amino acid sequences from plants are highly conserved. As with GluTR, two GSA-AT genes have been found in Arabidopsis and they may be differentially regulated by light. It has been reported that the presence of two genes for both enzymes of the C5 pathway indicate that there are two routes for ALA formation in chloroplasts (Kumar et al., Trends in Plant Science 1:371-376 (1996)). Transgenic tobacco plants that express antisense RNA to GSA-AT have been reported to show varying degrees of chlorophyll deficiency. Antisense plants with chlorophyll contents less than about 25% of that in the wild type plants which were maintained in the greenhouse under high light conditions, did not survive (Hennig et al., Proc. Natl. Acad. Sci. (U.S.A.) 94:4866-4871 (1997); Hoefgen, et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:1726-1730 (1994), both of which are herein incorporated by reference in their entirety).

[0013]The second phase of the biosynthesis of tetrapyrroles involves the formation of the porphyrin ring. The intermediates involved in this portion of the chlorophyll/heme biosynthetic pathway, from ALA to protoporphyrin IX, appear to be essentially the same in all organisms including plants and mammals.

[0014]Porphobilinogen synthase (E.C. 4.2.1.24), also known as ALA dehydratase, catalyzes the asymmetric condensation of two molecules of ALA to yield porphobilinogen (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). Porphobilinogen synthase is a metalloenzyme and there are different types of the enzyme categorized according to metal ion usage. Porphobilinogen synthase has been identified in several plants including spinach, pea, tomato, radish, and soybean. In higher plants the enzyme is located in the plastid, is a hexamer (40-50 kD subunits) and binds Mg⁺². The mammalian enzyme is an octamer and binds Zn²+ (Cheung et al., Biochemistry 36:1148-1156 (1997); Senior et al., Biochem. J. 320:401-412 (1996), both of which are herein incorporated by reference in their entirety). Several studies have shown that porphobilinogen synthase is both developmentally and light regulated in plants (Kyriacou et al., J. Am. Soc. Hortic. Sci. 121:91-95 (1996), the entirety of which is herein incorporated by reference in its entirety).

[0015]Hydroxymethylbilane synthase (E.C. 4.3.1.8), also known as porphobilinogen deaminase, catalyzes the formation of the linear tetrapyrrole hydroxymethylbilane (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). The reaction involves the deamination and polymerization of four molecules of the monopyrrole porphobilinogen. Hydroxymethylbilane synthase is unusual in that it contains a novel dipyrromethane cofactor at the active site, which is self-assembled by the apoenzyme and is covalently attached to an invariant cysteine. The enzyme has been identified in mammals, yeast, bacteria, and plants (e.g., pea, spinach, Arabidopsis). Hydroxymethylbilane synthase exists as a monomer with a molecular weight of 33-44 kD. Hydroxymethylbilane synthase from Arabidopsis has been cloned and found to be localized in the plastid in both roots and leaves (Witty et al., Planta 199:557-564 (1996), the entirety of which is herein incorporated by reference). The 3-dimensional structure of porphobilinogen deaminase from E. coli has been determined (Louie et al., Proteins: Struct., Funct., Genet. 25:48-78 (1996), the entirety of which is herein incorporated by reference).

[0016]Uroporphyrinogen III (co)synthase (E.C. 4.2.1.75) catalyzes the ring closure of the unstable linear tetrapyrrole hydroxymethylbilane and the simultaneous isomerization of the acetyl and propionyl groups at pyrrole ring D forming uroporphyrinogen III (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). Uroporphyrinogen III (co)synthase has been isolated from a number of sources including mammals, bacteria, and plants (spinach). Uroporphyrinogen III (co)synthase has a molecular weight of about 30 kD and is highly diverse in primary structure depending on the source.

[0017]Uroporphyrinogen III decarboxylase (E.C. 4.1.1.37) catalyzes the stepwise decarboxylation of all four acetate side chains of uroporphyrinogen III starting with ring D followed by rings A, B, and C, respectively, to form coproporphyrinogen III (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). At high substrate concentrations, decarboxylation can occur randomly. Uroporphyrinogen III decarboxylase has been isolated from mammals, yeast, bacteria and plants (e.g., tobacco, barley). It is a monomeric enzyme with a molecular weight of about 40 kD. The barley and tobacco enzymes are reported to be light regulated (Mock et al., Plant Mol. Biol. 28:245-256 (1995), the entirety of which is herein incorporated by reference). Antisense tobacco plants have been generated and decreased levels of the enzyme were accompanied by a light-dependent necrotic phenotype and accumulation of uroporphyrinogen. It has been reported that the lesions may be caused by reactive oxygen species generated by photooxidized uroporphyrinogen (Mock et al., Plant Mol. Biol. 28:245-256 (1995), the entirety of which is herein incorporated by reference).

[0018]In aerobic organisms including plants, coproporphyrinogen III oxidase (E.C. 1.3.3.3), catalyzes the oxygen dependent sequential oxidative decarboxylation of the A and B propionyl side chains of coproporphyrinogen III to yield two vinyl groups and protoporphyrinogen IX (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). A separate enzyme is reported to catalyze the anaerobic reaction.

[0019]Coproporphyrinogen III oxidase has been studied in a number of organisms including plants (tobacco, pea). The enzyme is a homodimer and has a subunit molecular weight of about 35-40 kD and is located in plastids. It has been reported that coproporphyrinogen III oxidase is peripherally associated with the membrane. It has been isolated from soybean, barley and tobacco and these sequences show 70% identity at the amino acid level. Transcript levels are reportedly similar in etiolated and green leaves (barley) but higher in developing cells than in mature cells (Kruse et al., Planta 196:796-803 (1995), the entirety of which is herein incorporated by reference). Antisense tobacco plants have been reported with decreased levels of the enzyme. The decreased level was accompanied by accumulation of coproporphyrinogen, slightly reduced chlorophyll content and a necrotic phenotype. The prominent phenotype indicates photodynamic damage (Kruse et al., EMBO J. 14:3712-3720 (1995), the entirety of which is herein incorporated by reference).

[0020]Protoporphyrinogen IX oxidase (E.C. 1.3.3.4) catalyzes the formation of the aromatic protoporphyrin IX by the six electron oxidation of protoporphyrinogen IX (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). This is the last reported common step in tetrapyrrole biosynthesis. In aerobic organisms, the reaction is catalyzed by a flavoprotein that utilizes oxygen as an oxidant and, under anaerobic conditions, the oxidation is achieved by passing electrons to the electron transport chain. The enzyme has been purified from a number of sources including mammals and plants (barley) and is an integral membrane protein. The barley enzyme has a molecular weight of 36 kD and activity has been found in both plastidal and mitochondrial extracts.

[0021]The plastidal and mitochondrial forms of protoporphyrinogen IX oxidase have been cloned from tobacco and were found to exhibit low homology. The mitochondrial form is associated with heme biosynthesis. The plastidic enzyme functions primarily in the formation of chlorophyll and to a lesser extent in the formation of heme required for plastid proteins (Lermontova et al., Proc. Natl. Acad. Sci. (U.S.A.) 94:8895-8900 (1997), the entirety of which is herein incorporated by reference). Protoporphyrinogen IX oxidase is susceptible to inhibition by a number of herbicides including diphenyl ethers. Phytotoxicity has been explained as due to the accumulation of excess protoporphyrinogen which is rapidly oxidized to protoporphyrin in the cytoplasm. Protoporphyrin has been reported as a potent photosensitizer which generates singlet oxygen and causes rapid lipid peroxidation and cell death.

[0022]In the third and final phase of tetrapyrrole biosynthesis, magnesium or iron is inserted into protoporphyrin IX and subsequent modifications lead to the synthesis of the final tetrapyrrole products, such as chlorophyll and heme.

[0023]Mg-chelatase catalyzes the conversion of protoporphyrin IX to magnesium protoporphyrin IX by the insertion of Mg⁺² (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). Mg-chelatase, which requires ATP, is reportedly a three component enzyme. The three protein components have molecular weights of about 140, 40, and 70 kD. The reaction takes place in two steps, an ATP-dependent activation followed by an ATP-dependent chelation step. Mg-chelatase activity has been demonstrated in peas, cucumber, and barley and reportedly is localized in the chloroplast. Barley, Arabidopsis, and soybean genes encoding the 140 and 40 kD subunits have been cloned. Studies with the two identified plant genes show that Mg-chelatase expression is light regulated (Walker and Willows, Biochem. J. 327:321-333 (1997), the entirety of which is herein incorporated by reference).

[0024]Mg-protoporphyrin IX O-methyltransferase (E.C. 2.1.1.11) esterifies the propionic side chain of ring III of Mg-protoporphyrin IX to form Mg-protoporphyrin IX monomethylester (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). The methyl group is donated by the cofactor S-adenosyl-L-methionine. The enzyme has been isolated from bacteria and plants (wheat). The gene for Mg-protoporphyrin IX O-methyltransferase has been cloned from bacteria including Synechocystis (Smith et al., Plant Mol. Biol. 30:1307-1314 (1996), the entirety of which is herein incorporated by reference).

[0025]Mg-protoporphyrin IX monomethyl ester cyclase catalyzes the cyclization of Mg-protoporphyrin IX monomethylester to form the isocyclic ring E of divinyl protochlorophyllide (Porra, Photochemistry and Photobiology 65:492-516 (1997)). In aerobic organisms the enzymatic reaction is dependent on O₂ and NADPH. Evidence suggests that Mg-protoporphyrin IX monomethyl ester cyclase is a membrane-bound monooxygenase of the iron-sulfur protein or copper protein type. Mg-protoporphyrin IX monomethyl ester cyclase has been extracted from chloroplasts of higher plants including cucumber and wheat. A cucumber enzyme has been shown to consist of two components, a soluble and a membrane-bound component. The soluble component has a molecular weight of 30 kD (Bollivar and Beale, Plant Physiol. 112:105-114 (1996), the entirety of which is herein incorporated by reference).

[0026]The reduction of divinyl protochlorophyllide to monovinyl protochlorophyllide has been reported based on product characterization, this reaction is catalyzed by 8-vinyl reductase (Porra, Photochemistry and Photobiology 65:492-516 (1997)). It has been reported that Mg-protoporphyrin IX monomethylester may also act as a substrate. NADPH is the most likely reductant. 8-vinyl reductase has been detected in higher plants including wheat and cucumber.

[0027]Protochlorophyllide reductase ("POR") (E.C. 1.3.1.33) catalyzes the reduction of the double bond between carbons 7 and 8 of the D ring of protochlorophyllide producing chlorophyllide (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). In angiosperms this is a light-dependent reaction. Non-flowering land plants, algae, and cyanobacteria contain both a light-dependent and a light-independent enzyme. Some other organisms contain only the light-independent enzyme. Three chloroplast genes have been identified that are essential for the light-independent enzyme (chlL, chlN and chlB).

[0028]The light-dependent POR ("L-POR") has been purified from barley, oat, and Arabidopsis. L-POR has a molecular weight of 35-38 kD and forms different multimers and aggregates with other proteins. L-POR is localized in the plastid and encoded in the nucleus. The genes encoding L-POR have been cloned from, for example, barley, Arabidopsis, pea, and oat. Two distinct and differentially light-regulated L-POR genes, POR A and POR B, have been identified in Arabidopsis and barley. POR A and POR B have biochemically equivalent light-dependent activities, with different expression patterns. POR B is reported to be present throughout the plant life cycle, while POR A is reported to function only in the very early stages of greening of etiolated tissue (Runge et al., Plant J. 9:513-523 (1996); Holtorf and Apel, Plant Mol. Biol. 31:387-392 (1996); Martin et al., Biochem. J. 325:139-145 (1997), all of which are herein incorporated by reference in their entirety).

[0029]Chlorophyll synthetase catalyzes the last reported step in chlorophyll a biosynthesis (Porra, Photochemistry and Photobiology 65:492-516 (1997); von Wettstein et al., Plant Cell 7:1039-1057 (1995)). Chlorophyll synthetase esterifies the propionic acid side chain of ring D of chlorophyllide with either phytyl pyrophosphate in green plants or geranylgeranyl pyrophosphate in greening etiolated seedlings. The enzyme is located in the plastid. A gene that encodes the enzyme in Synechocystis (chlG) and a gene that encodes the enzyme in Arabidopsis (G4) have been cloned and expressed in E. coli. The Synechocystis enzyme has the preferred substrate specificity reported for green plants. The cloned and expressed enzyme from Arabidopsis has the preferred substrate specificity reported for etiolated plants (Oster et al., J. Biol. Chem. 272:9671-9676 (1997); Oster and Rudiger, Bot. Acta 110:420-423 (1997), both of which are herein incorporated by reference).

[0030]Ferrochelatase (E.C. 4.99.1.1) catalyzes the conversion of protoporphyrin IX to heme. In plants the enzyme is located in both mitochondria and plastids. Ferrochelatase is reported to be a single soluble protein. Two ferrochelatase genes have been identified in Arabidopsis. Ferrochelatase-II encodes a protein targeted to the chloroplast and ferrochelatase-I encodes a protein targeted to both chloroplasts and mitochondria (Roper and Smith, Eur. J. Biochem. 246:32-37 (1997); Chow et al., J. Biol. Chem. 272:27565-27571 (1997), both of which are herein incorporated by reference).

[0031]II. Expressed Sequence Tag Nucleic Acid Molecules

[0032]Expressed sequence tags, or ESTs are randomly sequenced members of a cDNA library (or complementary DNA)(McCombie et al., Nature Genetics 1:124-130 (1992); Kurata et al., Nature Genetics 8:365-372 (1994); Okubo et al., Nature Genetics 2:173-179 (1992), all of which references are incorporated herein in their entirety). The randomly selected clones comprise insets that can represent a copy of up to the full length of a mRNA transcript.

[0033]Using conventional methodologies, cDNA libraries can be constructed from the mRNA (messenger RNA) of a given tissue or organism using poly dT primers and reverse transcriptase (Efstratiadis et al., Cell 7:279-3680 (1976), the entirety of which is herein incorporated by reference; Higuchi et al., Proc. Natl. Acad. Sci. (U.S.A.) 73:3146-3150 (1976), the entirety of which is herein incorporated by reference; Maniatis et al., Cell 8:163-182 (1976) the entirety of which is herein incorporated by reference; Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference; Okayama et al., Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference; Gubler et al., Gene 25:263-269 (1983), the entirety of which is herein incorporated by reference).

[0034]Several methods may be employed to obtain full-length cDNA constructs. For example, terminal transferase can be used to add homopolymeric tails of dC residues to the free 3' hydroxyl groups (Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference). This tail can then be hybridized by a poly dG oligo which can act as a primer for the synthesis of full length second strand cDNA. Okayama and Berg, Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference, report a method for obtaining full length cDNA constructs. This method has been simplified by using synthetic primer-adapters that have both homopolymeric tails for priming the synthesis of the first and second strands and restriction sites for cloning into plasmids (Coleclough et al., Gene 34:305-314 (1985), the entirety of which is herein incorporated by reference) and bacteriophage vectors (Krawinkel et al., Nucleic Acids Res. 14:1913 (1986), the entirety of which is herein incorporated by reference; Han et al., Nucleic Acids Res. 15:6304 (1987), the entirety of which is herein incorporated by reference).

[0035]These strategies have been coupled with additional strategies for isolating rare mRNA populations. For example, a typical mammalian cell contains between 10,000 and 30,000 different mRNA sequences (Davidson, Gene Activity in Early Development, 2nd ed., Academic Press, New York (1976), the entirety of which is herein incorporated by reference). The number of clones required to achieve a given probability that a low-abundance mRNA will be present in a cDNA library is N=(ln(1-P))/(ln(1-1/n)) where N is the number of clones required, P is the probability desired and 1/n is the fractional proportion of the total mRNA that is represented by a single rare mRNA (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press (1989), the entirety of which is herein incorporated by reference).

[0036]A method to enrich preparations of mRNA for sequences of interest is to fractionate by size. One such method is to fractionate by electrophoresis through an agarose gel (Pennica et al., Nature 301:214-221 (1983), the entirety of which is herein incorporated by reference). Another such method employs sucrose gradient centrifugation in the presence of an agent, such as methylmercuric hydroxide, that denatures secondary structure in RNA (Schweinfest et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:4997-5000 (1982), the entirety of which is herein incorporated by reference).

[0037]A frequently adopted method is to construct equalized or normalized cDNA libraries (Ko, Nucleic Acids Res. 18:5705-5711 (1990), the entirety of which is herein incorporated by reference; Patanjali et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:1943-1947 (1991), the entirety of which is herein incorporated by reference). Typically, the cDNA population is normalized by subtractive hybridization (Schmid et al., J. Neurochem. 48:307-312 (1987), the entirety of which is herein incorporated by reference; Fargnoli et al., Anal. Biochem. 187:364-373 (1990), the entirety of which is herein incorporated by reference; Travis et al., Proc. Natl. Acad. Sci (U.S.A.) 85:1696-1700 (1988), the entirety of which is herein incorporated by reference; Kato, Eur. J. Neurosci. 2:704-711 (1990); and Schweinfest et al., Genet. Anal. Tech. Appl. 7:64-70 (1990), the entirety of which is herein incorporated by reference). Subtraction represents another method for reducing the population of certain sequences in the cDNA library (Swaroop et al., Nucleic Acids Res. 19:1954 (1991), the entirety of which is herein incorporated by reference).

[0038]ESTs can be sequenced by a number of methods. Two basic methods may be used for DNA sequencing, the chain termination method of Sanger et al., Proc. Natl. Acad. Sci. (U.S.A.) 74:5463-5467 (1977), the entirety of which is herein incorporated by reference and the chemical degradation method of Maxam and Gilbert, Proc. Nat. Acad. Sci. (U.S.A.) 74:560-564 (1977), the entirety of which is herein incorporated by reference. Automation and advances in technology such as the replacement of radioisotopes with fluorescence-based sequencing have reduced the effort required to sequence DNA (Craxton, Methods 2:20-26 (1991), the entirety of which is herein incorporated by reference; Ju et al., Proc. Natl. Acad. Sci. (U.S.A.) 92:4347-4351 (1995), the entirety of which is herein incorporated by reference; Tabor and Richardson, Proc. Natl. Acad. Sci. (U.S.A.) 92:6339-6343 (1995), the entirety of which is herein incorporated by reference). Automated sequencers are available from, for example, Pharmacia Biotech, Inc., Piscataway, N.J. (Pharmacia ALF), LI-COR, Inc., Lincoln, Nebr. (LI-COR 4,000) and Millipore, Bedford, Mass. (Millipore BaseStation).

[0039]In addition, advances in capillary gel electrophoresis have also reduced the effort required to sequence DNA and such advances provide a rapid high resolution approach for sequencing DNA samples (Swerdlow and Gesteland, Nucleic Acids Res. 18:1415-1419 (1990); Smith, Nature 349:812-813 (1991); Luckey et al., Methods Enzymol. 218:154-172 (1993); Lu et al., J. Chromatog. A. 680:497-501 (1994); Carson et al., Anal. Chem. 65:3219-3226 (1993); Huang et al., Anal. Chem. 64:2149-2154 (1992); Kheterpal et al., Electrophoresis 17:1852-1859 (1996); Quesada and Zhang, Electrophoresis 17:1841-1851 (1996); Baba, Yakugaku Zasshi 117:265-281 (1997), all of which are herein incorporated by reference in their entirety).

[0040]ESTs longer than 150 nucleotides have been found to be useful for similarity searches and mapping (Adams et al., Science 252:1651-1656 (1991), herein incorporated by reference). ESTs, which can represent copies of up to the full length transcript, may be partially or completely sequenced. Between 150-450 nucleotides of sequence information is usually generated as this is the length of sequence information that is routinely and reliably produced using single run sequence data. Typically, only single run sequence data is obtained from the cDNA library (Adams et al., Science 252:1651-1656 (1991). Automated single run sequencing typically results in an approximately 2-3% error or base ambiguity rate (Boguski et al., Nature Genetics 4:332-333 (1993), the entirety of which is herein incorporated by reference).

[0041]EST databases have been constructed or partially constructed from, for example, C. elegans (McCombrie et al., Nature Genetics 1:124-131 (1992)), human liver cell line HepG2 (Okubo et al., Nature Genetics 2:173-179 (1992)), human brain RNA (Adams et al., Science 252:1651-1656 (1991); Adams et al., Nature 355:632-635 (1992)), Arabidopsis, (Newman et al., Plant Physiol. 106:1241-1255 (1994)); and rice (Kurata et al., Nature Genetics 8:365-372 (1994)).

[0042]III. Sequence Comparisons

[0043]A characteristic feature of a DNA sequence is that it can be compared with other DNA sequences. Sequence comparisons can be undertaken by determining the similarity of the test or query sequence with sequences in publicly available or proprietary databases ("similarity analysis") or by searching for certain motifs ("intrinsic sequence analysis")(e.g. cis elements)(Coulson, Trends in Biotechnology 12:76-80 (1994), the entirety of which is herein incorporated by reference); Birren et al., Genome Analysis 1: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997), the entirety of which is herein incorporated by reference).

[0044]Similarity analysis includes database search and alignment. Examples of public databases include the DNA Database of Japan (DDBJ)(on the Worldwide web at ddbj.nig.ac.jp/); Genebank (on the Worldwide web at ncbi.nlm.nih.gov/Web/Search/Index.htlm); and the European Molecular Biology Laboratory Nucleic Acid Sequence Database (EMBL) (on the Worldwide web at ebi.ac.uk/ebi_docs/embl_db/embl-db.html). Other appropriate databases include dbEST (on the Worldwide web at ncbi.nlm.nih.gov/dbEST/index.html), SwissProt (on the Worldwide web at ebi.ac.uk/ebi_docs/swisprot_db/swisshome.html), PIR (on the Worldwide web at nbrt.georgetown.edu/pir/) and The Institute for Genome Research (on the Worldwide web at tigr.org/tdb/tdb.html).

[0045]A number of different search algorithms have been developed, one example of which are the suite of programs referred to as BLAST programs. There are five implementations of BLAST, three designed for nucleotide sequences queries (BLASTN, BLASTX and TBLASTX) and two designed for protein sequence queries (BLASTP and TBLASTN) (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997)).

[0046]BLASTN takes a nucleotide sequence (the query sequence) and its reverse complement and searches them against a nucleotide sequence database. BLASTN was designed for speed, not maximum sensitivity and may not find distantly related coding sequences. BLASTX takes a nucleotide sequence, translates it in three forward reading frames and three reverse complement reading frames and then compares the six translations against a protein sequence database. BLASTX is useful for sensitive analysis of preliminary (single-pass) sequence data and is tolerant of sequencing errors (Gish and States, Nature Genetics 3:266-272 (1993), the entirety of which is herein incorporated by reference). BLASTN and BLASTX may be used in concert for analyzing EST data (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1:543-559 (1997)).

[0047]Given a coding nucleotide sequence and the protein it encodes, it is often preferable to use the protein as the query sequence to search a database because of the greatly increased sensitivity to detect more subtle relationships. This is due to the larger alphabet of proteins (20 amino acids) compared with the alphabet of nucleic acid sequences (4 bases), where it is far easier to obtain a match by chance. In addition, with nucleotide alignments, only a match (positive score) or a mismatch (negative score) is obtained, but with proteins, the presence of conservative amino acid substitutions can be taken into account. Here, a mismatch may yield a positive score if the non-identical residue has physical/chemical properties similar to the one it replaced. Various scoring matrices are used to supply the substitution scores of all possible amino acid pairs. A general purpose scoring system is the BLOSUM62 matrix (Henikoff and Henikoff, Proteins 17:49-61 (1993), the entirety of which is herein incorporated by reference), which is currently the default choice for BLAST programs. BLOSUM62 is tailored for alignments of moderately diverged sequences and thus may not yield the best results under all conditions. Altschul, J. Mol. Biol. 36:290-300 (1993), the entirety of which is herein incorporated by reference, describes a combination of three matrices to cover all contingencies. This may improve sensitivity, but at the expense of slower searches. In practice, a single BLOSUM62 matrix is often used but others (PAM40 and PAM250) may be attempted when additional analysis is necessary. Low PAM matrices are directed at detecting very strong but localized sequence similarities, whereas high PAM matrices are directed at detecting long but weak alignments between very distantly related sequences.

[0048]Homologues in other organisms are available that can be used for comparative sequence analysis. Multiple alignments are performed to study similarities and differences in a group of related sequences. CLUSTAL W is a multiple sequence alignment package that performs progressive multiple sequence alignments based on the method of Feng and Doolittle, J. Mol. Evol. 25:351-360 (1987), the entirety of which is herein incorporated by reference. Each pair of sequences is aligned and the distance between each pair is calculated; from this distance matrix, a guide tree is calculated and all of the sequences are progressively aligned based on this tree. A feature of the program is its sensitivity to the effect of gaps on the alignment; gap penalties are varied to encourage the insertion of gaps in probable loop regions instead of in the middle of structured regions. Users can specify gap penalties, choose between a number of scoring matrices, or supply their own scoring matrix for both pairwise alignments and multiple alignments. CLUSTAL W for UNIX and VMS systems is available at: ftp.ebi.ac.uk. Another program is MACAW (Schuler et al., Proteins Struct. Func. Genet. 9:180-190 (1991), the entirety of which is herein incorporated by reference, for which both Macintosh and Microsoft Windows versions are available. MACAW uses a graphical interface, provides a choice of several alignment algorithms and is available by anonymous ftp at: ncbi.nlm.nih.gov (directory/pub/macaw).

[0049]Sequence motifs are derived from multiple alignments and can be used to examine individual sequences or an entire database for subtle patterns. With motifs, it is sometimes possible to detect distant relationships that may not be demonstrable based on comparisons of primary sequences alone. Currently, the largest collection of sequence motifs in the world is PROSITE (Bairoch and Bucher, Nucleic Acid Research 22:3583-3589 (1994), the entirety of which is herein incorporated by reference). PROSITE may be accessed via either the ExPASy server on the World Wide Web or anonymous ftp site. Many commercial sequence analysis packages also provide search programs that use PROSITE data.

[0050]A resource for searching protein motifs is the BLOCKS E-mail server developed by Henikoff, Trends Biochem Sci. 18:267-268 (1993), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Nucleic Acid Research 19:6565-6572 (1991), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Proteins 17:49-61 (1993). BLOCKS searches a protein or nucleotide sequence against a database of protein motifs or "blocks." Blocks are defined as short, ungapped multiple alignments that represent highly conserved protein patterns. The blocks themselves are derived from entries in PROSITE as well as other sources. Either a protein query or a nucleotide query can be submitted to the BLOCKS server; if a nucleotide sequence is submitted, the sequence is translated in all six reading frames and motifs are sought for these conceptual translations. Once the search is completed, the server will return a ranked list of significant matches, along with an alignment of the query sequence to the matched BLOCKS entries.

[0051]Conserved protein domains can be represented by two-dimensional matrices, which measure either the frequency or probability of the occurrences of each amino acid residue and deletions or insertions in each position of the domain. This type of model, when used to search against protein databases, is sensitive and usually yields more accurate results than simple motif searches. Two popular implementations of this approach are profile searches such as GCG program ProfileSearch and Hidden Markov Models (HMMs)(Krough et al., J. Mol. Biol. 235:1501-1531, (1994); Eddy, Current Opinion in Structural Biology 6:361-365, (1996), both of which are herein incorporated by reference in their entirety). In both cases, a large number of common protein domains have been converted into profiles, as present in the PROSITE library, or HHM models, as in the Pfam protein domain library (Sonnhammer et al., Proteins 28:405-420 (1997), the entirety of which is herein incorporated by reference). Pfam contains more than 500 HMM models for enzymes, transcription factors, signal transduction molecules and structural proteins. Protein databases can be queried with these profiles or HMM models, which will identify proteins containing the domain of interest. For example, HMMSW or HMMFS, two programs in a public domain package called HMMER (Sonnhammer et al., Proteins 28:405-420 (1997)) can be used.

[0052]PROSITE and BLOCKS represent collected families of protein motifs. Thus, searching these databases entails submitting a single sequence to determine whether or not that sequence is similar to the members of an established family. Programs working in the opposite direction compare a collection of sequences with individual entries in the protein databases. An example of such a program is the Motif Search Tool, or MoST (Tatusov et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:12091-12095 (1994), the entirety of which is herein incorporated by reference). On the basis of an aligned set of input sequences, a weight matrix is calculated by using one of four methods (selected by the user). A weight matrix is simply a representation, position by position of how likely a particular amino acid will appear. The calculated weight matrix is then used to search the databases. To increase sensitivity, newly found sequences are added to the original data set, the weight matrix is recalculated and the search is performed again. This procedure continues until no new sequences are found.

SUMMARY OF THE INVENTION

[0053]The present invention provides a substantially purified nucleic acid molecule that encodes a maize or soybean tetrapyrrole pathway protein or fragment thereof, wherein the maize or soybean tetrapyrrole pathway protein is selected from the group consisting of: (a) putative chlorophyll synthetase enzyme; (b) protochlorophyllide reductase enzyme; (c) putative protochlorophyllide reductase enzyme; (d) coproporphyrinogen oxidase enzyme; (e) protoporphyrinogen oxidase enzyme; (f) uroporphyrinogen decarboxylase enzyme; (g) putative uroporphyrinogen decarboxylase enzyme (h) porphobilinogen synthase enzyme; (i) hydroxymethylbilane synthase enzyme; (j) glutamate-1-semialdehyde 2,1-aminomutase enzyme; (k) glutamate tRNA ligase enzyme; (l) glutamyl-tRNA reductase enzyme; (m) Mg-chelatase enzyme, and (n) ferrochelatase enzyme.

[0054]The present invention also provides a substantially purified nucleic acid molecule that encodes a plant tetrapyrrole pathway protein or fragment thereof, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or fragment thereof, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or fragment thereof

[0055]The present invention also provides a substantially purified maize or soybean tetrapyrrole pathway protein or fragment thereof, wherein the maize or soybean tetrapyrrole pathway protein is selected from the group consisting of (a) putative chlorophyll synthetase enzyme or fragment thereof; (b) putative protochlorophyllide reductase enzyme or fragment thereof; (c) protochlorophyllide reductase enzyme or fragment thereof; (d) coproporphyrinogen oxidase enzyme or fragment thereof; (e) protoporphyrinogen oxidase enzyme or fragment thereof; (f) uroporphyrinogen decarboxylase enzyme or fragment thereof; (g) putative uroporphyrinogen decarboxylase enzyme or fragment thereof; (h) porphobilinogen synthase enzyme or fragment thereof; (i) hydroxymethylbilane synthase enzyme or fragment thereof; (j) glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof; (k) glutamate tRNA ligase enzyme or fragment thereof; (l) glutamyl-tRNA reductase enzyme or fragment thereof; (m) Mg-chelatase enzyme or fragment thereof; and (n) ferrochelatase enzyme or fragment thereof.

[0056]The present invention also provides a substantially purified maize or soybean tetrapyrrole pathway protein or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 677.

[0057]The present invention also provides a substantially purified maize or soybean putative chlorophyll synthetase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 8 and SEQ ID NO: 384 through SEQ ID NO: 397.

[0058]The present invention also provides a substantially purified maize or soybean putative chlorophyll synthetase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 8 and SEQ ID NO: 384 through SEQ ID NO: 397.

[0059]The present invention also provides a substantially purified maize or soybean protochlorophyllide reductase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement SEQ ID NO: 9 through SEQ ID NO: 94 and SEQ ID NO: 398 through SEQ ID NO: 466.

[0060]The present invention also provides a substantially purified maize or soybean protochlorophyllide reductase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 9 through SEQ ID NO: 94 and SEQ ID NO: 398 through SEQ ID NO: 466.

[0061]The present invention also provides a substantially purified maize or soybean putative protochlorophyllide reductase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement SEQ ID NO: 95 through SEQ ID NO: 96 and SEQ ID NO: 467 through SEQ ID NO: 479.

[0062]The present invention also provides a substantially purified maize or soybean putative protochlorophyllide reductase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 95 through SEQ ID NO: 96 and SEQ ID NO: 467 through SEQ ID NO: 479.

[0063]The present invention also provides a substantially purified maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence consisting of a complement of SEQ ID NO: 97 through SEQ ID NO: 128 and SEQ ID NO: 480 through SEQ ID NO: 494.

[0064]The present invention also provides a substantially purified maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof encoded by a nucleic acid sequence consisting of SEQ ID NO: 97 through SEQ ID NO: 128 and SEQ ID NO: 480 through SEQ ID NO: 494.

[0065]The present invention also provides a substantially purified maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 129 through SEQ ID NO: 131 and SEQ ID NO: 495 through SEQ ID NO: 499.

[0066]The present invention also provides a substantially purified maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 129 through SEQ ID NO: 131 and SEQ ID NO: 495 through SEQ ID NO: 499.

[0067]The present invention also provides a substantially purified maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 132 through SEQ ID NO: 144 and SEQ ID NO: 500 through SEQ ID NO: 509.

[0068]The present invention also provides a substantially purified maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 132 through SEQ ID NO: 144 and SEQ ID NO: 500 through SEQ ID NO: 509.

[0069]The present invention also provides a substantially purified a maize putative uroporphyrinogen decarboxylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence consisting of a complement of SEQ ID NO: 510.

[0070]The present invention also provides a substantially purified maize putative uroporphyrinogen decarboxylase enzyme or fragment thereof encoded by a nucleic acid sequence consisting of SEQ ID NO: 510.

[0071]The present invention also provides a substantially purified soybean porphobilinogen synthetase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 145 through SEQ ID NO: 191 and SEQ ID NO: 511 through SEQ ID NO: 531.

[0072]The present invention also provides a substantially purified maize or soybean porphobilinogen synthetase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 145 through SEQ ID NO: 191 and SEQ ID NO: 511 through SEQ ID NO: 531.

[0073]The present invention also provides a substantially purified maize or soybean hydroxymethylbilane synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement SEQ ID NO: 154, SEQ ID NO: 192 through SEQ ID NO: 217 and SEQ ID NO: 532 through SEQ ID NO: 542.

[0074]The present invention also provides a substantially purified maize or soybean hydroxymethylbilane enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 154, SEQ ID NO: 192 through SEQ ID NO: 217 and SEQ ID NO: 532 through SEQ ID NO: 542.

[0075]The present invention also provides a substantially purified maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence consisting of a complement of SEQ ID NO: 218 through SEQ ID NO: 265 and SEQ ID NO: 543 through SEQ ID NO: 569.

[0076]The present invention also provides a substantially purified maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof encoded by a nucleic acid sequence consisting of SEQ ID NO: 218 through SEQ ID NO: 265 and SEQ ID NO: 543 through SEQ ID NO: 569.

[0077]The present invention also provides a substantially purified maize or soybean glutamate tRNA ligase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585.

[0078]The present invention also provides a substantially purified maize or soybean glutamate tRNA ligase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585.

[0079]The present invention also provides a substantially purified maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585.

[0080]The present invention also provides a substantially purified maize or soybean glutamyl-tRNA reductase enzyme fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585.

[0081]The present invention also provides a substantially purified maize or soybean Mg-chelatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 290 through SEQ ID NO: 306 and SEQ ID NO: 586 through SEQ ID NO: 609.

[0082]The present invention also provides a substantially purified maize or soybean Mg-chelatase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 290 through SEQ ID NO: 306 and SEQ ID NO: 586 through SEQ ID NO: 609.

[0083]The present invention also provides a substantially purified maize or soybean ferrochelatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 372 through SEQ ID NO: 383 and SEQ ID NO: 653 through SEQ ID NO:677.

[0084]The present invention also provides a substantially purified maize or soybean ferrochelatase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 372 through SEQ ID NO: 383 and SEQ ID NO: 653 through SEQ ID NO:677.

[0085]The present invention also provides a purified antibody or fragment thereof which is capable of specifically binding to a maize or soybean tetrapyrrole pathway protein or fragment thereof, wherein the maize or soybean tetrapyrrole pathway protein or fragment thereof is encoded by a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of consisting of SEQ ID NO: 1 through SEQ ID NO: 677.

[0086]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a putative maize or soybean chlorophyll synthetase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 8 and SEQ ID NO: 384 through SEQ ID NO: 397 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 1 through SEQ ID NO: 8 and SEQ ID NO: 384 through SEQ ID NO: 397.

[0087]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean protochlorophyllide reductase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 9 through SEQ ID NO: 94 and SEQ ID NO: 398 through SEQ ID NO: 466 or a nucleic acid sequence selected from the group consisting of SEQ ID NO: 95 through SEQ ID NO: 96 and SEQ ID NO: 398 through SEQ ID NO: 466.

[0088]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean putative protochlorophyllide reductase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 95 through SEQ ID NO: 96 and SEQ ID NO: 467 through SEQ ID NO: 479 or a nucleic acid sequence selected from the group consisting of SEQ ID NO: 95 through SEQ ID NO: 96 and SEQ ID NO: 467 through SEQ ID NO: 479.

[0089]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean coproporphyrinogen oxidase or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule consisting of a compliment of a nucleic acid sequence having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 97 through SEQ ID NO: 128 and SEQ ID NO: 480 through SEQ ID NO: 494 or a nucleic acid sequence selected from the group consisting of SEQ ID NO: 97 through SEQ ID NO: 128 and SEQ ID NO: 480 through SEQ ID NO: 494.

[0090]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 129 through SEQ ID NO: 131 and SEQ ID NO: 495 through SEQ ID NO: 499 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 129 through SEQ ID NO: 131 and SEQ ID NO: 495 through SEQ ID NO: 499.

[0091]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 132 through SEQ ID NO: 144 and SEQ ID NO: 500 through SEQ ID NO: 509 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 132 through SEQ ID NO: 144 and SEQ ID NO: 500 through SEQ ID NO: 509.

[0092]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence consisting of a complement of SEQ ID NO: 510 or a nucleic acid sequence consisting SEQ ID NO: 510.

[0093]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean porphobilinogen enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 145 through SEQ ID NO: 191 and SEQ ID NO: 511 through SEQ ID NO: 531 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 145 through SEQ ID NO: 191 and SEQ ID NO: 511 through SEQ ID NO: 531.

[0094]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean hydroxymethylbilane synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 154, SEQ ID NO: 192 through SEQ ID NO: 217 and SEQ ID NO: 532 through SEQ ID NO: 542 or a nucleic acid sequence selected from the group consisting of SEQ ID NO: 154, SEQ ID NO: 192 through SEQ ID NO: 217 and SEQ ID NO: 532 through SEQ ID NO: 542.

[0095]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule consisting of a compliment of a nucleic acid sequence having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 218 through SEQ ID NO: 265 and SEQ ID NO: 543 through SEQ ID NO: 569 or a nucleic acid sequence selected from the group consisting of SEQ ID NO: 218 through SEQ ID NO: 265 and SEQ ID NO: 543 through SEQ ID NO: 569.

[0096]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean glutamate tRNA ligase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585.

[0097]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 290 through SEQ ID NO: 306 and SEQ ID NO: 586 through SEQ ID NO: 609 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 290 through SEQ ID NO: 306 and SEQ ID NO: 586 through SEQ ID NO: 609.

[0098]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean Mg-chelatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 307 through SEQ ID NO: 371 and SEQ ID NO: 610 through SEQ ID NO: 652 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 307 through SEQ ID NO: 371 and SEQ ID NO: 610 through SEQ ID NO: 652.

[0099]The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or soybean ferrochelatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 372 through SEQ ID NO: 383 and SEQ ID NO: 653 through SEQ ID NO: 677 or a nucleic acid sequence selected from the group consisting SEQ ID NO: 372 through SEQ ID NO: 383 and SEQ ID NO: 653 through SEQ ID NO: 677.

[0100]The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; (B) a structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (a) a nucleic acid sequence which encodes for a putative chlorophyll synthetase enzyme or fragment thereof; (b) a nucleic acid sequence which encodes for a protochlorophyllide reductase or fragment thereof; (c) a nucleic acid sequence which encodes for a putative protochlorophyllide reductase or fragment thereof; (d) a nucleic acid sequence which encodes for a coproporphyrinogen oxidase or fragment thereof; (e) a nucleic acid sequence which encodes for a protoporphyrinogen oxidase enzyme or fragment thereof; (f) a nucleic acid sequence which encodes for a uroporphyrinogen decarboxylase enzyme or fragment thereof; (g) a nucleic acid sequence which encodes for a putative uroporphyrinogen decarboxylase enzyme or fragment thereof; (h) a nucleic acid sequence which encodes for a porphobilinogen synthase enzyme or fragment thereof; (i) a nucleic acid sequence which encodes for a hydroxymethylbilane synthase enzyme or fragment thereof; (j) a nucleic acid sequence which encodes for a glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof; (k) a nucleic acid sequence which encodes for a glutamate tRNA ligase enzyme or fragment thereof; (l) a nucleic acid sequence which encodes for a glutamyl-tRNA reductase enzyme or fragment thereof; (m) a nucleic acid sequence which encodes for a Mg-chelatase enzyme or fragment thereof; and (n) a nucleic acid sequence which encodes for a ferrochelatase enzyme or fragment thereof (m) a nucleic acid sequence which is complementary to any of the nucleic acid sequences of (a) through (n); and (C) a 3' non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of the mRNA molecule.

[0101]The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule encodes a plant tetrapyrrole pathway protein or fragment thereof, the structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof; which is linked to (C) a 3' non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of the mRNA molecule.

[0102]The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or fragment thereof, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean coproporphyrinogen oxidase or fragment thereof, a nucleic acid sequence which encodes a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof, a nucleic acid sequence which encodes a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean porphobilinogen synthase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean hydroxymethylbilane synthase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean glutamate tRNA ligase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof, a nucleic acid sequence which encodes a maize or soybean Mg-chelatase enzyme or fragment thereof, and a nucleic acid sequence which encodes a maize or soybean ferrochelatase enzyme or fragment thereof; which is linked to (C) a 3' non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of the mRNA molecule.

[0103]The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof; which is linked to (C) a 3' non-translated sequence that functions in plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of the mRNA molecule.

[0104]The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to: (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to an endogenous mRNA molecule having a nucleic acid sequence selected from the group consisting of an endogenous mRNA molecule that encodes a putative maize or soybean putative chlorophyll synthetase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean porphobilinogen synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean hydromethylbilane synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or fragment thereof an endogenous mRNA molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean Mg-chelatase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or soybean ferrochelatase enzyme or fragment thereof; which is linked to (C) a 3' non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of the mRNA molecule.

[0105]The present invention also provides a method for determining a level or pattern of a plant tetrapyrrole pathway protein in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant tetrapyrrole pathway protein; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant tetrapyrrole pathway protein.

[0106]The present invention also provides a method for determining a level or pattern of a plant tetrapyrrole pathway protein in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant tetrapyrrole pathway protein; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant tetrapyrrole pathway protein.

[0107]The present invention also provides a method for determining a level or pattern of a plant tetrapyrrole pathway protein in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant tetrapyrrole pathway protein, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or reference plant tissue with the known level or pattern of the plant tetrapyrrole pathway protein.

[0108]The present invention also provides a method for determining a level or pattern of a plant tetrapyrrole pathway protein in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant tetrapyrrole pathway protein, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or the reference plant tissue with the known level or pattern of the plant tetrapyrrole pathway protein.

[0109]The present invention provides a method of determining a mutation in a plant whose presence is predictive of a mutation affecting a level or pattern of a protein comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid, the marker nucleic acid selected from the group of marker nucleic acid molecules which specifically hybridize to a nucleic acid molecule having a nucleic acid sequence selected from the group of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragment of either and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the protein in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

[0110]The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant tetrapyrrole pathway protein comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant tetrapyrrole pathway protein in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

[0111]The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant tetrapyrrole pathway protein comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant tetrapyrrole pathway protein in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

[0112]The present invention also provides a method of producing a plant containing an overexpressed protein comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region has a nucleic acid sequence selected from group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 wherein the structural region is linked to a 3' non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the protein; and (B) growing the transformed plant.

[0113]The present invention also provides a method of producing a plant containing an overexpressed plant tetrapyrrole pathway protein comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof; wherein the structural region is linked to a 3' non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant tetrapyrrole pathway protein; and (B) growing the transformed plant.

[0114]The present invention also provides a method of producing a plant containing an overexpressed plant tetrapyrrole pathway protein comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof or fragment and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof or fragment of either, wherein the structural region is linked to a 3' non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant tetrapyrrole pathway protein; and (B) growing the transformed plant.

[0115]The present invention also provides a method of producing a plant containing reduced levels of a plant tetrapyrrole pathway protein comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677; wherein the structural region is linked to a 3' non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant tetrapyrrole pathway protein; and (B) growing the transformed plant.

[0116]The present invention also provides a method of producing a plant containing reduced levels of a plant tetrapyrrole pathway protein comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof or fragment, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof or fragment and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof or fragment of either, wherein the structural region is linked to a 3' non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant tetrapyrrole pathway protein; and (B) growing the transformed plant.

[0117]The present invention also provides a method for reducing expression of a plant tetrapyrrole pathway protein in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragments of either and the transcribed strand is complementary to an endogenous mRNA molecule; and wherein the transcribed nucleic acid molecule is linked to a 3' non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and (B) growing the transformed plant.

[0118]The present invention also provides a method for reducing expression of a plant tetrapyrrole pathway protein in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to a nucleic acid molecule selected from the group consisting of an endogenous mRNA molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean porphobilinogen synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean hydromethylbilane synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or soybean Mg-chelatase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or soybean ferrochelatase enzyme or fragment thereof, and wherein the transcribed nucleic acid molecule is linked to a 3' non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and (B) growing the transformed plant.

[0119]The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule has a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragment of either; and (B) calculating the degree of association between the polymorphism and the plant trait.

[0120]The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof or fragment of either and (B) calculating the degree of association between the polymorphism and the plant trait.

[0121]The present invention also provides a method of isolating a nucleic acid that encodes a plant tetrapyrrole pathway protein or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragment of either with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the first nucleic acid molecule and the second nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

[0122]The present invention also provides a method of isolating a nucleic acid molecule that encodes a plant tetrapyrrole pathway protein or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof or fragment of either, with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the plant tetrapyrrole pathway protein nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

DETAILED DESCRIPTION OF THE INVENTION

Definitions and Agents of the Present Invention

Definitions:

[0123]As used herein, a tetrapyrrole pathway enzyme is any molecule that is associated with the biosynthesis or degradation of tetrapyrroles.

[0124]As used herein, ALA refers to 5-aminolevunic acid and 4-aminolevulinate.

[0125]As used herein, ALA synthase (E.C. 2.3.1.37) refers to any enzyme that catalyzes the condensation of glycine with succinyl-CoA to generate ALA.

[0126]As used herein, glutamyl-tRNA synthetase (GluRS) (E.C. 6.1.1.17) refers to any enzyme that converts glutamate to glutamyl-tRNA (Glu-tRNA).

[0127]As used herein, glutamyl-tRNA reductase (GluTR) refers to any enzyme that catalyzes the NADPH dependent reduction of Glu-tRNA to glutamate 1-semialdehyde (GSA) with the release of intact tRNA.

[0128]As used herein, glutamate-1-semialdehyde aminotransferase (GSA-AT) (E.C. 5.4.3.8) refers to any enzyme that catalyzes the conversion of GSA to ALA

[0129]As used herein, porphobilinogen synthase (ALA dehydratase) (E.C. 4.2.1.24) refers to any enzyme that catalyzes the asymmetric condensation of two molecules of ALA to yield porphobilinogen.

[0130]As used herein, porphobilinogen deaminase (hydroxymethylbilane synthase) (E.C. 4.3.1.8) refers to any enzyme that catalyzes the formation of the linear tetrapyrrole hydroxymethylbilane.

[0131]As used herein, uroporphyrinogen III (co)synthase (E.C. 4.2.1.75) refers to any enzyme that catalyzes the ring closure of the unstable linear tetrapyrrole hydroxymethylbilane and the simultaneous isomerization of the acetyl and propionyl groups at pyrrole ring D forming uroporphyrinogen III.

[0132]As used herein, uroporphyrinogen III decarboxylase (E.C. 4.1.1.37) refers to any enzyme that catalyzes the stepwise decarboxylation of all four acetate side chains of uroporphyrinogen III starting with ring D followed by rings A, B, and C respectively to form coproporphyrinogen III.

[0133]As used herein, coproporphyrinogen III oxidase (E.C. 1.3.3.3) refers to any enzyme that catalyzes the oxygen dependent sequential oxidative decarboxylation of the A and B propionyl side chains of coproporphyrinogen III to yield two vinyl groups and protoporphyrinogen IX.

[0134]As used herein, protoporphyrinogen IX oxidase (E.C. 1.3.3.4) refers to any enzyme that catalyzes the formation of the aromatic protoporphyrin IX by the six electron oxidation of protoporphyrinogen IX.

[0135]As used herein, Mg-chelatase refers to any enzyme that catalyzes the conversion of protoporphyrin IX to magnesium protoporphyrin IX by the insertion Mg⁺².

[0136]As used herein, Mg-protoporphyrin IX O-methyltransferase (E.C. 2.1.1.11) refers to any enzyme that esterifies the propionic side chain of ring III of Mg-protoporphyrin IX to form Mg-protoporphyrin IX monomethylester.

[0137]As used herein, Mg-protoporphyrin IX monomethyl ester cyclase refers to any enzyme that catalyzes the cyclization of Mg-protoporphyrin IX monomethylester to form the isocyclic ring E of divinyl protochlorophyllide.

[0138]As used herein, 8-vinyl reductase refers to any enzyme that can reduce divinyl protochlorophyllide or Mg-protoporphyrin IX monomethylester to monovinyl protochlorophyllide.

[0139]As used herein, protochlorophyllide reductase ("POR") (E.C. 1.3.1.33) refers to any enzyme that catalyzes the reduction of the double bond between carbons 7 and 8 of the D ring of protochlorophyllide producing chlorophyllide

[0140]As used herein, chlorophyll synthetase refers to any enzyme that esterifies the propionic acid side chain of ring D of chlorophyllide with either phytyl pyrophosphate or geranylgeranyl pyrophosphate.

[0141]As used herein, ferrochelatase (E.C. 4.99.1.1) refers to any enzyme that catalyzes the conversion of protoporphyrin IX to heme.

Agents

[0142](a) Nucleic Acid Molecules

[0143]Agents of the present invention include plant nucleic acid molecules and more preferably include maize, soybean and Arabidopsis thaliana nucleic acid molecules and more preferably include nucleic acid molecules of the maize genotypes B73 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), B73×Mo17 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), DK604 (Dekalb Genetics, Dekalb, Ill. U.S.A.), H99 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), RX601 (Asgrow Seed Company, Des Moines, Iowa), Mo17 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), and soybean types Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa), C1944 (United States Department of Agriculture (USDA) Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), FT108 (Monsoy, Brazil), Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), BW211S Null (Tohoku University, Morioka, Japan), PI507354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Asgrow A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.), PI227687 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.).

[0144]A subset of the nucleic acid molecules of the present invention includes nucleic acid molecules that are marker molecules. Another subset of the nucleic acid molecules of the present invention include nucleic acid molecules that encode a protein or fragment thereof. Another subset of the nucleic acid molecules of the present invention are EST molecules.

[0145]Fragment nucleic acid molecules may encode significant portion(s) of, or indeed most of, these nucleic acid molecules. Alternatively, the fragments may comprise smaller oligonucleotides (having from about 15 to about 250 nucleotide residues and more preferably, about 15 to about 30 nucleotide residues).

[0146]As used herein, an agent, be it a naturally occurring molecule or otherwise may be "substantially purified," if desired, such that one or more molecules that is or may be present in a naturally occurring preparation containing that molecule will have been removed or will be present at a lower concentration than that at which it would normally be found.

[0147]The agents of the present invention will preferably be "biologically active" with respect to either a structural attribute, such as the capacity of a nucleic acid to hybridize to another nucleic acid molecule, or the ability of a protein to be bound by an antibody (or to compete with another molecule for such binding). Alternatively, such an attribute may be catalytic and thus involve the capacity of the agent to mediate a chemical reaction or response.

[0148]The agents of the present invention may also be recombinant. As used herein, the term recombinant means any agent (e.g. DNA, peptide etc.), that is, or results, however indirect, from human manipulation of a nucleic acid molecule.

[0149]It is understood that the agents of the present invention may be labeled with reagents that facilitate detection of the agent (e.g. fluorescent labels, Prober et al., Science 238:336-340 (1987); Albarella et al., EP 144914; chemical labels, Sheldon et al., U.S. Pat. No. 4,582,789; Albarella et al., U.S. Pat. No. 4,563,417; modified bases, Miyoshi et al., EP 119448, all of which are hereby incorporated by reference in their entirety).

[0150]It is further understood, that the present invention provides recombinant bacterial, mammalian, microbial, insect, fungal and plant cells and viral constructs comprising the agents of the present invention. (See, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells)

[0151]Nucleic acid molecules or fragments thereof of the present invention are capable of specifically hybridizing to other nucleic acid molecules under certain circumstances. As used herein, two nucleic acid molecules are said to be capable of specifically hybridizing to one another if the two molecules are capable of forming an anti-parallel, double-stranded nucleic acid structure. A nucleic acid molecule is said to be the "complement" of another nucleic acid molecule if they exhibit complete complementarity. As used herein, molecules are said to exhibit "complete complementarity" when every nucleotide of one of the molecules is complementary to a nucleotide of the other. Two molecules are said to be "minimally complementary" if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under at least conventional "low-stringency" conditions. Similarly, the molecules are said to be "complementary" if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional "high-stringency" conditions. Conventional stringency conditions are described by Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd Ed., Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989) and by Haymes et al., Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington, D.C. (1985), the entirety of which is herein incorporated by reference. Departures from complete complementarity are therefore permissible, as long as such departures do not completely preclude the capacity of the molecules to form a double-stranded structure. Thus, in order for a nucleic acid molecule to serve as a primer or probe it need only be sufficiently complementary in sequence to be able to form a stable double-stranded structure under the particular solvent and salt concentrations employed.

[0152]Appropriate stringency conditions which promote DNA hybridization, for example, 6.0×sodium chloride/sodium citrate (SSC) at about 45° C., followed by a wash of 2.0×SSC at 50° C., are known to those skilled in the art or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the salt concentration in the wash step can be selected from a low stringency of about 2.0×SSC at 50° C. to a high stringency of about 0.2×SSC at 50° C. In addition, the temperature in the wash step can be increased from low stringency conditions at room temperature, about 22° C., to high stringency conditions at about 65° C. Both temperature and salt may be varied, or either the temperature or the salt concentration may be held constant while the other variable is changed.

[0153]In a preferred embodiment, a nucleic acid of the present invention will specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof under moderately stringent conditions, for example at about 2.0×SSC and about 65° C.

[0154]In a particularly preferred embodiment, a nucleic acid of the present invention will include those nucleic acid molecules that specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof under high stringency conditions such as 0.2×SSC and about 65° C.

[0155]In one aspect of the present invention, the nucleic acid molecules of the present invention have one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof. In another aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 90% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof. In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 95% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof. In a more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 98% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof. In an even more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 99% sequence identity with one or more of the sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof.

[0156]In a further more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention exhibit 100% sequence identity with a nucleic acid molecule present within MONN01, SATMON001, SATMON003 through SATMON014, SATMON016 through SATMON031, SATMON033, SATMON034, SATMON˜001, SATMONN01, SATMONN04 through SATMONN006, CMz029 through CMz031, CMz033 through CMz037, CMz039 through CMz042, CMz044 through CMz045, CMz047 through CMz050, SOYMON001 through SOYMON038, Soy51 through Soy56, Soy58 through Soy62, Soy65 through Soy73 and Soy76 through Soy77, Lib9, Lib22 through Lib25, Lib35, and Lib146 (Monsanto Company, St. Louis, Mo. U.S.A.).

[0157](i) Nucleic Acid Molecules Encoding Proteins or Fragments Thereof

[0158]Nucleic acid molecules of the present invention can comprise sequences that encode a tetrapyrrole pathway enzyme or fragment thereof. Such transcription factors or fragments thereof include homologues of known transcription factors in other organisms.

[0159]In a preferred embodiment of the present invention, a maize or soybean tetrapyrrole pathway enzyme or fragment thereof of the present invention is a homologue of another plant tetrapyrrole pathway protein. In another preferred embodiment of the present invention, a maize or soybean tetrapyrrole pathway enzyme or fragment thereof of the present invention is a homologue of a fungal tetrapyrrole pathway enzyme. In another preferred embodiment of the present invention, a maize or soybean tetrapyrrole pathway enzyme of the present invention is a homologue of mammalian transcription factor. In another preferred embodiment of the present invention, a maize or soybean tetrapyrrole pathway enzyme or fragment thereof of the present invention is a homologue of a bacterial transcription factor. In another preferred embodiment of the present invention, a maize or soybean tetrapyrrole pathway enzyme or fragment thereof of the present invention is a homologue of a maize tetrapyrrole pathway enzyme. In another preferred embodiment of the present invention, a maize or soybean tetrapyrrole pathway enzyme homologue or fragment thereof of the present invention is a homologue of a soybean transcription factor.

[0160]In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or soybean tetrapyrrole pathway enzyme or fragment thereof where a maize or soybean tetrapyrrole pathway enzyme exhibits a BLAST probability score of greater than 1E-12, preferably a BLAST probability score of between about 1E-30 and about 1E-12, even more preferably a BLAST probability score of greater than 1E-30 with its homologue.

[0161]In another preferred embodiment of the present invention, the nucleic acid molecule encoding a maize or soybean tetrapyrrole pathway enzyme or fragment thereof exhibits a % identity with its homologue of between about 25% and about 40%, more preferably of between about 40 and about 70%, even more preferably of between about 70% and about 90% and even more preferably between about 90% and 99%. In another preferred embodiment, of the present invention, a maize or soybean tetrapyrrole enzyme or fragment thereof exhibits a % identity with its homologue of 100%.

[0162]In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or soybean tetrapyrrole pathway enzyme or fragment thereof where a maize or soybean tetrapyrrole pathway enzyme exhibits a BLAST score of greater than 120, preferably a BLAST score of between about 1450 and about 120, even more preferably a BLAST score of greater than 1450 with its homologue.

[0163]Nucleic acid molecules of the present invention also include non-maize, non-soybean homologues. Preferred non-homologues are selected from the group consisting of alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm and Phaseolus.

[0164]In a preferred embodiment, nucleic acid molecules having SEQ ID NO: 1 through SEQ ID NO: 677 or complements and fragments of either can be utilized to obtain such homologues.

[0165]The degeneracy of the genetic code, which allows different nucleic acid sequences to code for the same protein or peptide, is known in the literature. (U.S. Pat. No. 4,757,006, the entirety of which is herein incorporated by reference).

[0166]In an aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or soybean tetrapyrrole pathway enzyme or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 677 due to the degeneracy in the genetic code in that they encode the same transcription factor but differ in nucleic acid sequence.

[0167]In another further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or soybean tetrapyrrole pathway enzyme or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 677 due to fact that the different nucleic acid sequence encodes a transcription factor having one or more conservative amino acid residue. Examples of conservative substitutions are set forth in Table 1. It is understood that codons capable of coding for such conservative substitutions are known in the art.

TABLE-US-00001 TABLE 1 Original Residue Conservative Substitutions Ala Ser Arg Lys Asn Gln; His Asp Glu Cys Ser; Ala Gln Asn Glu Asp Gly Pro His Asn; Gln Ile Leu; Val Leu Ile; Val Lys Arg; Gln; Glu Met Leu; Ile Phe Met; Leu; Tyr Ser Thr Thr Ser Trp Tyr Tyr Trp; Phe Val Ile; Leu

[0168]In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or soybean tetrapyrrole or fragment thereof set forth in SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof due to the fact that one or more codons encoding an amino acid has been substituted for a codon that encodes a nonessential substitution of the amino acid originally encoded.

[0169]Agents of the present invention include nucleic acid molecules that encode a maize, or soybean tetrapyrrole pathway enzyme or fragment thereof and particularly substantially purified nucleic acid molecules selected from the group consisting of a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or fragment, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme fragment, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or fragment, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or fragment and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or fragment.

[0170]Non-limiting examples of such nucleic acid molecules of the present invention are nucleic acid molecules comprising: SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof that encode for a plant tetrapyrrole pathway protein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 8 and SEQ ID NO: 384 through SEQ ID NO: 397 or fragment thereof that encode for a putative chlorophyll synthetase enzyme or fragment thereof, SEQ ID NO: 9 through SEQ ID NO: 94 and SEQ ID NO: 398 through SEQ ID NO: 466 or fragment thereof that encode for a protochlorophyllide reductase enzyme or fragment thereof, SEQ ID NO: 95 through SEQ ID NO: 96 and SEQ ID NO: 467 through SEQ ID NO: 479 or fragment thereof that encode for a putative protochlorophyllide reductase enzyme or fragment thereof, SEQ ID NO: 97 through SEQ ID NO: 128 and SEQ ID NO: 480 through SEQ ID NO: 494 or fragment thereof that encodes for a coproporphyrinogen oxidase enzyme or fragment thereof, SEQ ID NO: 129 through SEQ ID NO: 131 and SEQ ID NO: 495 through SEQ ID NO: 499 or fragment thereof that encode for a protoporphyrinogen oxidase enzyme or fragment thereof, SEQ ID NO: 132 through SEQ ID NO: 144 and SEQ ID NO: 500 through SEQ ID NO: 509 or fragment thereof that encode for an uroporphyrinogen decarboxylase enzyme or fragment thereof, SEQ ID NO: 510 or fragment thereof that encode for a putative uroporphyrinogen decarboxylase enzyme or fragment thereof, SEQ ID NO: 145 through SEQ ID NO: 191 and SEQ ID NO: 511 through SEQ ID NO: 531 or fragment thereof that encode for a porphobilinogen synthase enzyme or fragment thereof, SEQ ID NO: 154, SEQ ID NO: 192 through SEQ ID NO: 217 and SEQ ID NO: 532 through SEQ ID NO: 542 or fragment thereof that encode for a hydroxymethylbilane synthase enzyme or fragment thereof, SEQ ID NO: 218 through SEQ ID NO: 265 and SEQ ID NO: 543 through SEQ ID NO: 569 or fragment thereof that encodes for a glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585 or fragment thereof that encode for a glutamate tRNA ligase enzyme or fragment thereof, SEQ ID NO: 290 through SEQ ID NO: 306 and SEQ ID NO: 586 through SEQ ID NO: 609 or fragment thereof that encode for an glutamyl-tRNA reductase enzyme or fragment thereof, SEQ ID NO: 307 through SEQ ID NO: 371 and SEQ ID NO: 610 through SEQ ID NO: 652 or fragment thereof that encode for a Mg-chelatase enzyme or fragment thereof, and SEQ ID NO: 372 through SEQ ID NO: 383 and SEQ ID NO: 653 through SEQ ID NO: 677 or fragment thereof that encode for an ferrochelatase enzyme or fragment thereof.

[0171]A nucleic acid molecule of the present invention can also encode an homologue of a putative maize or soybean chlorophyll synthetase enzyme or fragment thereof, a maize or soybean protochlorophyllide reductase enzyme or fragment or fragment thereof, a putative maize or soybean protochlorophyllide reductase enzyme or fragment or fragment thereof, a maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof, a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof, a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof, a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof, a maize or soybean porphobilinogen synthase enzyme or fragment thereof, a maize or soybean hydroxymethylbilane synthase enzyme or fragment thereof, a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, a maize or soybean glutamate tRNA ligase enzyme fragment thereof, a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof, a maize or soybean Mg-chelatase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or fragment thereof. As used herein a homologue protein molecule or fragment thereof is a counterpart protein molecule or fragment thereof in a second species (e.g., maize chlorophyll synthetase is a homologue of Arabidopsis' chlorophyll synthetase).

[0172](ii) Nucleic Acid Molecule Markers and Probes

[0173]One aspect of the present invention concerns markers that include nucleic acid molecules SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragments of either that can act as markers or other nucleic acid molecules of the present invention that can act as markers. Genetic markers of the present invention include "dominant" or "codominant" markers "Codominant markers" reveal the presence of two or more alleles (two per diploid individual) at a locus. "Dominant markers" reveal the presence of only a single allele per locus. The presence of the dominant marker phenotype (e.g., a band of DNA) is an indication that one allele is present in either the homozygous or heterozygous condition. The absence of the dominant marker phenotype (e.g. absence of a DNA band) is merely evidence that "some other" undefined allele is present. In the case of populations where individuals are predominantly homozygous and loci are predominately dimorphic, dominant and codominant markers can be equally valuable. As populations become more heterozygous and multi-allelic, codominant markers often become more informative of the genotype than dominant markers. Marker molecules can be, for example, capable of detecting polymorphisms such as single nucleotide polymorphisms (SNPs).

[0174]SNPs are single base changes in genomic DNA sequence. They occur at greater frequency and are spaced with a greater uniformly throughout a genome than other reported forms of polymorphism. The greater frequency and uniformity of SNPs means that there is greater probability that such a polymorphism will be found near or in a genetic locus of interest than would be the case for other polymorphisms. SNPs are located in protein-coding regions and noncoding regions of a genome. Some of these SNPs may result in defective or variant protein expression (e.g., as a results of mutations or defective splicing). Analysis (genotyping) of characterized SNPs can require only a plus/minus assay rather than a lengthy measurement, permitting easier automation.

[0175]SNPs can be characterized using any of a variety of methods. Such methods include the direct or indirect sequencing of the site, the use of restriction enzymes (Botstein et al., Am. J. Hum. Genet. 32:314-331 (1980), the entirety of which is herein incorporated reference; Konieczny and Ausubel, Plant J. 4:403-410 (1993), the entirety of which is herein incorporated by reference), enzymatic and chemical mismatch assays (Myers et al., Nature 313:495-498 (1985), the entirety of which is herein incorporated by reference), allele-specific PCR (Newton et al., Nucl. Acids Res. 17:2503-2516 (1989), the entirety of which is herein incorporated by reference; Wu et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:2757-2760 (1989), the entirety of which is herein incorporated by reference), ligase chain reaction (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is herein incorporated by reference), single-strand conformation polymorphism analysis (Labrune et al., Am. J. Hum. Genet. 48: 1115-1120 (1991), the entirety of which is herein incorporated by reference), primer-directed nucleotide incorporation assays (Kuppuswami et al., Proc. Natl. Acad. Sci. USA 88:1143-1147 (1991), the entirety of which is herein incorporated by reference), dideoxy fingerprinting (Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference), solid-phase ELISA-based oligonucleotide ligation assays (Nikiforov et al., Nucl. Acids Res. 22:4167-4175 (1994), the entirety of which is herein incorporated by reference), oligonucleotide fluorescence-quenching assays (Livak et al., PCR Methods Appl. 4:357-362 (1995), the entirety of which is herein incorporated by reference), 5'-nuclease allele-specific hybridization TaqMan assay (Livak et al., Nature Genet. 9:341-342 (1995), the entirety of which is herein incorporated by reference), template-directed dye-terminator incorporation (TDI) assay (Chen and Kwok, Nucl. Acids Res. 25:347-353 (1997), the entirety of which is herein incorporated by reference), allele-specific molecular beacon assay (Tyagi et al., Nature Biotech. 16: 49-53 (1998), the entirety of which is herein incorporated by reference), PinPoint assay (Haff and Smirnov, Genome Res. 7: 378-388 (1997), the entirety of which is herein incorporated by reference) and dCAPS analysis (Neff et al., Plant J. 14:387-392 (1998), the entirety of which is herein incorporated by reference).

[0176]Additional markers, such as AFLP markers, RFLP markers and RAPD markers, can be utilized (Walton, Seed World 22-29 (July, 1993), the entirety of which is herein incorporated by reference; Burow and Blake, Molecular Dissection of Complex Traits, 13-29, Paterson (ed.), CRC Press, New York (1988), the entirety of which is herein incorporated by reference). DNA markers can be developed from nucleic acid molecules using restriction endonucleases, the PCR and/or DNA sequence information. RFLP markers result from single base changes or insertions/deletions. These codominant markers are highly abundant in plant genomes, have a medium level of polymorphism and are developed by a combination of restriction endonuclease digestion and Southern blotting hybridization. CAPS are similarly developed from restriction nuclease digestion but only of specific PCR products. These markers are also codominant, have a medium level of polymorphism and are highly abundant in the genome. The CAPS result from single base changes and insertions/deletions.

[0177]Another marker type, RAPDs, are developed from DNA amplification with random primers and result from single base changes and insertions/deletions in plant genomes. They are dominant markers with a medium level of polymorphisms and are highly abundant. AFLP markers require using the PCR on a subset of restriction fragments from extended adapter primers. These markers are both dominant and codominant are highly abundant in genomes and exhibit a medium level of polymorphism.

[0178]SSRs require DNA sequence information. These codominant markers result from repeat length changes, are highly polymorphic and do not exhibit as high a degree of abundance in the genome as CAPS, AFLPs and RAPDs SNPs also require DNA sequence information. These codominant markers result from single base substitutions. They are highly abundant and exhibit a medium of polymorphism (Rafalski et al., In: Nonmammalian Genomic Analysis, Birren and Lai (ed.), Academic Press, San Diego, Calif., pp. 75-134 (1996), the entirety of which is herein incorporated by reference). It is understood that a nucleic acid molecule of the present invention may be used as a marker.

[0179]A PCR probe is a nucleic acid molecule capable of initiating a polymerase activity while in a double-stranded structure to with another nucleic acid. Various methods for determining the structure of PCR probes and PCR techniques exist in the art. Computer generated searches using programs such as Primer3 (on the Worldwide web at genome.wi.mit.edu/cgi-bin/primer/primer3.cgi), STSPipeline (on the Worldwide web at genome.wi.mit.edu/cgi-bin/www-STS_Pipeline), or GeneUp (Pesole et al., BioTechniques 25:112-123 (1998) the entirety of which is herein incorporated by reference), for example, can be used to identify potential PCR primers.

[0180]It is understood that a fragment of one or more of the nucleic acid molecules of the present invention may be a probe and specifically a PCR probe.

[0181](b) Protein and Peptide Molecules

[0182]A class of agents comprises one or more of the protein or fragments thereof or peptide molecules encoded by SEQ ID NO: 1 through SEQ ID NO: 677 or one or more of the protein or fragment thereof and peptide molecules encoded by other nucleic acid agents of the present invention. As used herein, the term "protein molecule" or "peptide molecule" includes any molecule that comprises five or more amino acids. It is well known in the art that proteins may undergo modification, including post-translational modifications, such as, but not limited to, disulfide bond formation, glycosylation, phosphorylation, or oligomerization. Thus, as used herein, the term "protein molecule" or "peptide molecule" includes any protein molecule that is modified by any biological or non-biological process. The terms "amino acid" and "amino acids" refer to all naturally occurring L-amino acids. This definition is meant to include norleucine, omithine, homocysteine and homoserine.

[0183]Non-limiting examples of the protein or fragment thereof of the present invention include a maize or soybean tetrapyrrole pathway enzyme or fragment thereof, a putative maize or soybean chlorophyll synthetase enzyme or fragment thereof, a maize or soybean protochlorophyllide reductase enzyme or fragment or fragment thereof, a putative maize or soybean protochlorophyllide reductase enzyme or fragment or fragment thereof, a maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof, a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof, a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof, a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof, a maize or soybean porphobilinogen synthase enzyme or fragment thereof, a maize or soybean hydroxymethylbilane synthase enzyme or fragment thereof, a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, a maize or soybean glutamate tRNA ligase enzyme fragment thereof, a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof, a maize or soybean Mg-chelatase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or fragment thereof.

[0184]Non-limiting examples of the protein or fragment molecules of the present invention are a transcription factor or fragment thereof encoded by: SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof that encode for a tetrapyrrole pathway enzyme or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 8 and SEQ ID NO: 384 through SEQ ID NO: 397 or fragment thereof that encode for a putative chlorophyll synthetase enzyme or fragment thereof, SEQ ID NO: 9 through SEQ ID NO: 94 and SEQ ID NO: 398 through SEQ ID NO: 466 or fragment thereof that encode for a protochlorophyllide reductase enzyme or fragment thereof, SEQ ID NO: 95 through SEQ ID NO: 96 and SEQ ID NO: 467 through SEQ ID NO: 479 or fragment thereof that encode for a putative protochlorophyllide reductase enzyme or fragment thereof, SEQ ID NO: 97 through SEQ ID NO: 128 and SEQ ID NO: 480 through SEQ ID NO: 494 or fragment thereof that encodes for a coproporphyrinogen oxidase enzyme or fragment thereof, SEQ ID NO: 129 through SEQ ID NO: 131 and SEQ ID NO: 495 through SEQ ID NO: 499 or fragment thereof that encode for a protoporphyrinogen oxidase enzyme or fragment thereof, SEQ ID NO: 132 through SEQ ID NO: 144 and SEQ ID NO: 500 through SEQ ID NO: 509 or fragment thereof that encode for an uroporphyrinogen decarboxylase enzyme or fragment thereof, SEQ ID NO: 510 or fragment thereof that encode for a putative uroporphyrinogen decarboxylase enzyme or fragment thereof, SEQ ID NO: 145 through SEQ ID NO: 191 and SEQ ID NO: 511 through SEQ ID NO: 531 or fragment thereof that encode for a porphobilinogen synthase enzyme or fragment thereof, SEQ ID NO: 154, SEQ ID NO: 192 through SEQ ID NO: 217 and SEQ ID NO: 532 through SEQ ID NO: 542 or fragment thereof that encode for a hydroxymethylbilane synthase enzyme or fragment thereof, SEQ ID NO: 218 through SEQ ID NO: 265 and SEQ ID NO: 543 through SEQ ID NO: 569 or fragment thereof that encodes for a glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, SEQ ID NO: 266 through SEQ ID NO: 289 and SEQ ID NO: 570 through SEQ ID NO: 585 or fragment thereof that encode for a glutamate tRNA ligase enzyme or fragment thereof, SEQ ID NO: 290 through SEQ ID NO: 306 and SEQ ID NO: 586 through SEQ ID NO: 609 or fragment thereof that encode for an glutamyl-tRNA reductase enzyme or fragment thereof, SEQ ID NO: 307 through SEQ ID NO: 371 and SEQ ID NO: 610 through SEQ ID NO: 652 or fragment thereof that encode for a Mg-chelatase enzyme or fragment thereof, and SEQ ID NO: 372 through SEQ ID NO: 383 and SEQ ID NO: 653 through SEQ ID NO: 677 or fragment thereof that encode for an ferrochelatase enzyme or fragment thereof.

[0185]One or more of the protein or fragment of peptide molecules may be produced via chemical synthesis, or more preferably, by expressing in a suitable bacterial or eucaryotic host. Suitable methods for expression are described by Sambrook et al., (In: Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989)), or similar texts. For example, the protein may be expressed in, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells.

[0186]A "protein fragment" is a peptide or polypeptide molecule whose amino acid sequence comprises a subset of the amino acid sequence of that protein. A protein or fragment thereof that comprises one or more additional peptide regions not derived from that protein is a "fusion" protein. Such molecules may be derivatized to contain carbohydrate or other moieties (such as keyhole limpet hemocyanin, etc.). Fusion protein or peptide molecules of the present invention are preferably produced via recombinant means.

[0187]Another class of agents comprise protein or peptide molecules or fragments or fusions thereof encoded by SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof in which conservative, non-essential or non-relevant amino acid residues have been added, replaced or deleted. Computerized means for designing modifications in protein structure are known in the art (Dahiyat and Mayo, Science 278:82-87 (1997), the entirety of which is herein incorporated by reference).

[0188]The protein molecules of the present invention include plant homologue proteins. An example of such a homologue is a homologue protein of a non-maize or non soybean plant species, that include but not limited to alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus etc. Particularly preferred non-maize or non-soybean for use for the isolation of homologs would include, Arabidopsis, barley, cotton, oat, oilseed rape, rice, canola, ornamentals, sugarcane, sugarbeet, tomato, potato, wheat and turf grasses. Such a homologue can be obtained by any of a variety of methods. Most preferably, as indicated above, one or more of the disclosed sequences (SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof) will be used to define a pair of primers that may be used to isolate the homologue-encoding nucleic acid molecules from any desired species. Such molecules can be expressed to yield homologues by recombinant means.

[0189](c) Antibodies

[0190]One aspect of the present invention concerns antibodies, single-chain antigen binding molecules, or other proteins that specifically bind to one or more of the protein or peptide molecules of the present invention and their homologues, fusions or fragments. Such antibodies may be used to quantitatively or qualitatively detect the protein or peptide molecules of the present invention. As used herein, an antibody or peptide is said to "specifically bind" to a protein or peptide molecule of the present invention if such binding is not competitively inhibited by the presence of non-related molecules.

[0191]Nucleic acid molecules that encode all or part of the protein of the present invention can be expressed, via recombinant means, to yield protein or peptides that can in turn be used to elicit antibodies that are capable of binding the expressed protein or peptide. Such antibodies may be used in immunoassays for that protein. Such protein-encoding molecules, or their fragments may be a "fusion" molecule (i.e., a part of a larger nucleic acid molecule) such that, upon expression, a fusion protein is produced. It is understood that any of the nucleic acid molecules of the present invention may be expressed, via recombinant means, to yield proteins or peptides encoded by these nucleic acid molecules.

[0192]The antibodies that specifically bind proteins and protein fragments of the present invention may be polyclonal or monoclonal and may comprise intact immunoglobulins, or antigen binding portions of immunoglobulins fragments (such as (F(ab'), F(ab')₂), or single-chain immunoglobulins producible, for example, via recombinant means. It is understood that practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of antibodies (see, for example, Harlow and Lane, In: Antibodies: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988), the entirety of which is herein incorporated by reference).

[0193]Murine monoclonal antibodies are particularly preferred. BALB/c mice are preferred for this purpose, however, equivalent strains may also be used. The animals are preferably immunized with approximately 25 μg of purified protein (or fragment thereof) that has been emulsified in a suitable adjuvant (such as TiterMax adjuvant (Vaxcel, Norcross, Ga.)). Immunization is preferably conducted at two intramuscular sites, one intraperitoneal site and one subcutaneous site at the base of the tail. An additional i.v. injection of approximately 25 μg of antigen is preferably given in normal saline three weeks later. After approximately 11 days following the second injection, the mice may be bled and the blood screened for the presence of anti-protein or peptide antibodies. Preferably, a direct binding Enzyme-Linked Immunoassay (ELISA) is employed for this purpose.

[0194]More preferably, the mouse having the highest antibody titer is given a third i.v. injection of approximately 25 μg of the same protein or fragment. The splenic leukocytes from this animal may be recovered 3 days later and then permitted to fuse, most preferably, using polyethylene glycol, with cells of a suitable myeloma cell line (such as, for example, the P3X63Ag8.653 myeloma cell line). Hybridoma cells are selected by culturing the cells under "HAT" (hypoxanthine-aminopterin-thymine) selection for about one week. The resulting clones may then be screened for their capacity to produce monoclonal antibodies ("mAbs"), preferably by direct ELISA.

[0195]In one embodiment, anti-protein or peptide monoclonal antibodies are isolated using a fusion of a protein or peptide of the present invention, or conjugate of a protein or peptide of the present invention, as immunogens. Thus, for example, a group of mice can be immunized using a fusion protein emulsified in Freund's complete adjuvant (e.g. approximately 50 μg of antigen per immunization). At three week intervals, an identical amount of antigen is emulsified in Freund's incomplete adjuvant and used to immunize the animals. Ten days following the third immunization, serum samples are taken and evaluated for the presence of antibody. If antibody titers are too low, a fourth booster can be employed. Polysera capable of binding the protein or peptide can also be obtained using this method.

[0196]In a preferred procedure for obtaining monoclonal antibodies, the spleens of the above-described immunized mice are removed, disrupted and immune splenocytes are isolated over a ficoll gradient. The isolated splenocytes are fused, using polyethylene glycol with BALB/c-derived HGPRT (hypoxanthine guanine phosphoribosyl transferase) deficient P3x63xAg8.653 plasmacytoma cells. The fused cells are plated into 96 well microtiter plates and screened for hybridoma fusion cells by their capacity to grow in culture medium supplemented with hypothanthine, aminopterin and thymidine for approximately 2-3 weeks.

[0197]Hybridoma cells that arise from such incubation are preferably screened for their capacity to produce an immunoglobulin that binds to a protein of interest. An indirect ELISA may be used for this purpose. In brief, the supernatants of hybridomas are incubated in microtiter wells that contain immobilized protein. After washing, the titer of bound immunoglobulin can be determined using, for example, a goat anti-mouse antibody conjugated to horseradish peroxidase. After additional washing, the amount of immobilized enzyme is determined (for example through the use of a chromogenic substrate). Such screening is performed as quickly as possible after the identification of the hybridoma in order to ensure that a desired clone is not overgrown by non-secreting neighbor cells. Desirably, the fusion plates are screened several times since the rates of hybridoma growth vary. In a preferred sub-embodiment, a different antigenic form may be used to screen the hybridoma. Thus, for example, the splenocytes may be immunized with one immunogen, but the resulting hybridomas can be screened using a different immunogen. It is understood that any of the protein or peptide molecules of the present invention may be used to raise antibodies.

[0198]As discussed below, such antibody molecules or their fragments may be used for diagnostic purposes. Where the antibodies are intended for diagnostic purposes, it may be desirable to derivatize them, for example with a ligand group (such as biotin) or a detectable marker group (such as a fluorescent group, a radioisotope or an enzyme).

[0199]The ability to produce antibodies that bind the protein or peptide molecules of the present invention permits the identification of mimetic compounds of those molecules. A "mimetic compound" is a compound that is not that compound, or a fragment of that compound, but which nonetheless exhibits an ability to specifically bind to antibodies directed against that compound.

[0200]It is understood that any of the agents of the present invention can be substantially purified and/or be biologically active and/or recombinant.

Uses of the Agents of the Invention

[0201]Nucleic acid molecules and fragments thereof of the present invention may be employed to obtain other nucleic acid molecules from the same species (e.g., ESTs or fragment thereof from maize may be utilized to obtain other nucleic acid molecules from maize). Such nucleic acid molecules include the nucleic acid molecules that encode the complete coding sequence of a protein and promoters and flanking sequences of such molecules. In addition, such nucleic acid molecules include nucleic acid molecules that encode for other isozymes or gene family members. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from maize or soybean. Methods for forming such libraries are well known in the art.

[0202]Nucleic acid molecules and fragments thereof of the present invention may also be employed to obtain nucleic acid homologues. Such homologues include the nucleic acid molecule of other plants or other organisms (e.g., alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus, etc.) including the nucleic acid molecules that encode, in whole or in part, protein homologues of other plant species or other organisms, sequences of genetic elements such as promoters and transcriptional regulatory elements. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from such plant species. Methods for forming such libraries are well known in the art. Such homologue molecules may differ in their nucleotide sequences from those found in one or more of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof because complete complementarity is not needed for stable hybridization. The nucleic acid molecules of the present invention therefore also include molecules that, although capable of specifically hybridizing with the nucleic acid molecules may lack "complete complementarity."

[0203]Any of a variety of methods may be used to obtain one or more of the above-described nucleic acid molecules (Zamechik et al., Proc. Natl. Acad. Sci. (U.S.A.) 83:4143-4146 (1986), the entirety of which is herein incorporated by reference; Goodchild et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:5507-5511 (1988), the entirety of which is herein incorporated by reference; Wickstrom et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:1028-1032 (1988), the entirety of which is herein incorporated by reference; Holt et al., Molec. Cell. Biol. 8:963-973 (1988), the entirety of which is herein incorporated by reference; Gerwirtz et al., Science 242:1303-1306 (1988), the entirety of which is herein incorporated by reference; Anfossi et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:3379-3383 (1989), the entirety of which is herein incorporated by reference; Becker et al., EMBO J. 8:3685-3691 (1989); the entirety of which is herein incorporated by reference). Automated nucleic acid synthesizers may be employed for this purpose. In lieu of such synthesis, the disclosed nucleic acid molecules may be used to define a pair of primers that can be used with the polymerase chain reaction (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al., European Patent 50,424; European Patent 84,796; European Patent 258,017; European Patent 237,362; Mullis, European Patent 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194, all of which are herein incorporated by reference in their entirety) to amplify and obtain any desired nucleic acid molecule or fragment.

[0204]Promoter sequence(s) and other genetic elements, including but not limited to transcriptional regulatory flanking sequences, associated with one or more of the disclosed nucleic acid sequences can also be obtained using the disclosed nucleic acid sequence provided herein. In one embodiment, such sequences are obtained by incubating EST nucleic acid molecules or preferably fragments thereof with members of genomic libraries (e.g. maize and soybean) and recovering clones that hybridize to the EST nucleic acid molecule or fragment thereof. In a second embodiment, methods of "chromosome walking," or inverse PCR may be used to obtain such sequences (Frohman et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8998-9002 (1988); Ohara et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:5673-5677 (1989); Pang et al., Biotechniques 22:1046-1048 (1977); Huang et al., Methods Mol. Biol. 69:89-96 (1997); Huang et al., Method Mol. Biol. 67:287-294 (1997); Benkel et al., Genet. Anal. 13:123-127 (1996); Hartl et al., Methods Mol. Biol. 58:293-301 (1996), all of which are herein incorporated by reference in their entirety).

[0205]The nucleic acid molecules of the present invention may be used to isolate promoters of cell enhanced, cell specific, tissue enhanced, tissue specific, developmentally or environmentally regulated expression profiles. Isolation and functional analysis of the 5' flanking promoter sequences of these genes from genomic libraries, for example, using genomic screening methods and PCR techniques would result in the isolation of useful promoters and transcriptional regulatory elements. These methods are known to those of skill in the art and have been described (See, for example, Birren et al., Genome Analysis: Analyzing DNA, 1, (1997), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference). Promoters obtained utilizing the nucleic acid molecules of the present invention could also be modified to affect their control characteristics. Examples of such modifications would include but are not limited to enhanced sequences as reported in Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants. Such genetic elements could be used to enhance gene expression of new and existing traits for crop improvements.

[0206]In one sub-aspect, such an analysis is conducted by determining the presence and/or identity of polymorphism(s) by one or more of the nucleic acid molecules of the present invention and more preferably one or more of the EST nucleic acid molecule or fragment thereof which are associated with a phenotype, or a predisposition to that phenotype.

[0207]Any of a variety of molecules can be used to identify such polymorphism(s). In one embodiment, one or more of the EST nucleic acid molecules (or a sub-fragment thereof) may be employed as a marker nucleic acid molecule to identify such polymorphism(s). Alternatively, such polymorphisms can be detected through the use of a marker nucleic acid molecule or a marker protein that is genetically linked to (i.e., a polynucleotide that co-segregates with) such polymorphism(s).

[0208]In an alternative embodiment, such polymorphisms can be detected through the use of a marker nucleic acid molecule that is physically linked to such polymorphism(s). For this purpose, marker nucleic acid molecules comprising a nucleotide sequence of a polynucleotide located within 1 mb of the polymorphism(s) and more preferably within 100 kb of the polymorphism(s) and most preferably within 10 kb of the polymorphism(s) can be employed.

[0209]The genomes of animals and plants naturally undergo spontaneous mutation in the course of their continuing evolution (Gusella, Ann. Rev. Biochem. 55:831-854 (1986)). A "polymorphism" is a variation or difference in the sequence of the gene or its flanking regions that arises in some of the members of a species. The variant sequence and the "original" sequence co-exist in the species' population. In some instances, such co-existence is in stable or quasi-stable equilibrium.

[0210]A polymorphism is thus said to be "allelic," in that, due to the existence of the polymorphism, some members of a species may have the original sequence (i.e., the original "allele") whereas other members may have the variant sequence (i.e., the variant "allele"). In the simplest case, only one variant sequence may exist and the polymorphism is thus said to be di-allelic. In other cases, the species' population may contain multiple alleles and the polymorphism is termed tri-allelic, etc. A single gene may have multiple different unrelated polymorphisms. For example, it may have a di-allelic polymorphism at one site and a multi-allelic polymorphism at another site.

[0211]The variation that defines the polymorphism may range from a single nucleotide variation to the insertion or deletion of extended regions within a gene. In some cases, the DNA sequence variations are in regions of the genome that are characterized by short tandem repeats (STRs) that include tandem di- or tri-nucleotide repeated motifs of nucleotides. Polymorphisms characterized by such tandem repeats are referred to as "variable number tandem repeat" ("VNTR") polymorphisms. VNTRs have been used in identity analysis (Weber, U.S. Pat. No. 5,075,217; Armour et al., FEBS Lett. 307:113-115 (1992); Jones et al., Eur. J. Haematol. 39:144-147 (1987); Horn et al., PCT Patent Application WO91/14003; Jeffreys, European Patent Application 370,719; Jeffreys, U.S. Pat. No. 5,175,082; Jeffreys et al., Amer. J. Hum. Genet. 39:11-24 (1986); Jeffreys et al., Nature 316:76-79 (1985); Gray et al., Proc. R. Acad. Soc. Lond. 243:241-253 (1991); Moore et al., Genomics 10:654-660 (1991); Jeffreys et al., Anim. Genet. 18:1-15 (1987); Hillel et al., Anim. Genet. 20:145-155 (1989); Hillel et al., Genet. 124:783-789 (1990), all of which are herein incorporated by reference in their entirety).

[0212]The detection of polymorphic sites in a sample of DNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis or other means.

[0213]The most preferred method of achieving such amplification employs the polymerase chain reaction ("PCR") (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al., European Patent Appln. 50,424; European Patent Appln. 84,796; European Patent Application 258,017; European Patent Appln. 237,362; Mullis, European Patent Appln. 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194), using primer pairs that are capable of hybridizing to the proximal sequences that define a polymorphism in its double-stranded form.

[0214]In lieu of PCR, alternative methods, such as the "Ligase Chain Reaction" ("LCR") may be used (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is herein incorporated by reference). LCR uses two pairs of oligonucleotide probes to exponentially amplify a specific target. The sequences of each pair of oligonucleotides is selected to permit the pair to hybridize to abutting sequences of the same strand of the target. Such hybridization forms a substrate for a template-dependent ligase. As with PCR, the resulting products thus serve as a template in subsequent cycles and an exponential amplification of the desired sequence is obtained.

[0215]LCR can be performed with oligonucleotides having the proximal and distal sequences of the same strand of a polymorphic site. In one embodiment, either oligonucleotide will be designed to include the actual polymorphic site of the polymorphism. In such an embodiment, the reaction conditions are selected such that the oligonucleotides can be ligated together only if the target molecule either contains or lacks the specific nucleotide that is complementary to the polymorphic site present on the oligonucleotide. Alternatively, the oligonucleotides may be selected such that they do not include the polymorphic site (see, Segev, PCT Application WO 90/01069, the entirety of which is herein incorporated by reference).

[0216]The "Oligonucleotide Ligation Assay" ("OLA") may alternatively be employed (Landegren et al., Science 241:1077-1080 (1988), the entirety of which is herein incorporated by reference). The OLA protocol uses two oligonucleotides which are designed to be capable of hybridizing to abutting sequences of a single strand of a target. OLA, like LCR, is particularly suited for the detection of point mutations. Unlike LCR, however, OLA results in "linear" rather than exponential amplification of the target sequence.

[0217]Nickerson et al., have described a nucleic acid detection assay that combines attributes of PCR and OLA (Nickerson et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8923-8927 (1990), the entirety of which is herein incorporated by reference). In this method, PCR is used to achieve the exponential amplification of target DNA, which is then detected using OLA. In addition to requiring multiple and separate, processing steps, one problem associated with such combinations is that they inherit all of the problems associated with PCR and OLA.

[0218]Schemes based on ligation of two (or more) oligonucleotides in the presence of nucleic acid having the sequence of the resulting "di-oligonucleotide", thereby amplifying the di-oligonucleotide, are also known (Wu et al., Genomics 4:560-569 (1989), the entirety of which is herein incorporated by reference) and may be readily adapted to the purposes of the present invention.

[0219]Other known nucleic acid amplification procedures, such as allele-specific oligomers, branched DNA technology, transcription-based amplification systems, or isothermal amplification methods may also be used to amplify and analyze such polymorphisms (Malek et al., U.S. Pat. No. 5,130,238; Davey et al., European Patent Application 329,822; Schuster et al., U.S. Pat. No. 5,169,766; Miller et al., PCT Patent Application WO 89/06700; Kwoh et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1173-1177 (1989); Gingeras et al., PCT Patent Application WO 88/10315; Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:392-396 (1992), all of which are herein incorporated by reference in their entirety).

[0220]The identification of a polymorphism can be determined in a variety of ways. By correlating the presence or absence of it in a plant with the presence or absence of a phenotype, it is possible to predict the phenotype of that plant. If a polymorphism creates or destroys a restriction endonuclease cleavage site, or if it results in the loss or insertion of DNA (e.g., a VNTR polymorphism), it will alter the size or profile of the DNA fragments that are generated by digestion with that restriction endonuclease. As such, individuals that possess a variant sequence can be distinguished from those having the original sequence by restriction fragment analysis. Polymorphisms that can be identified in this manner are termed "restriction fragment length polymorphisms" ("RFLPs"). RFLPs have been widely used in human and plant genetic analyses (Glassberg, UK Patent Application 2135774; Skolnick et al., Cytogen. Cell Genet. 32:58-67 (1982); Botstein et al., Ann. J. Hum. Genet. 32:314-331 (1980); Fischer et al., (PCT Application WO90/13668); Uhlen, PCT Application WO90/11369).

[0221]Polymorphisms can also be identified by Single Strand Conformation Polymorphism (SSCP) analysis. SSCP is a method capable of identifying most sequence variations in a single strand of DNA, typically between 150 and 250 nucleotides in length (Elles, Methods in Molecular Medicine: Molecular Diagnosis of Genetic Diseases, Humana Press (1996), the entirety of which is herein incorporated by reference); Orita et al., Genomics 5:874-879 (1989), the entirety of which is herein incorporated by reference). Under denaturing conditions a single strand of DNA will adopt a conformation that is uniquely dependent on its sequence conformation. This conformation usually will be different, even if only a single base is changed. Most conformations have been reported to alter the physical configuration or size sufficiently to be detectable by electrophoresis. A number of protocols have been described for SSCP including, but not limited to, Lee et al., Anal. Biochem. 205:289-293 (1992), the entirety of which is herein incorporated by reference; Suzuki et al., Anal. Biochem. 192:82-84 (1991), the entirety of which is herein incorporated by reference; Lo et al., Nucleic Acids Research 20:1005-1009 (1992), the entirety of which is herein incorporated by reference; Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference. It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by SSCP analysis.

[0222]Polymorphisms may also be found using a DNA fingerprinting technique called amplified fragment length polymorphism (AFLP), which is based on the selective PCR amplification of restriction fragments from a total digest of genomic DNA to profile that DNA (Vos et al., Nucleic Acids Res. 23:4407-4414 (1995), the entirety of which is herein incorporated by reference). This method allows for the specific co-amplification of high numbers of restriction fragments, which can be visualized by PCR without knowledge of the nucleic acid sequence.

[0223]AFLP employs basically three steps. Initially, a sample of genomic DNA is cut with restriction enzymes and oligonucleotide adapters are ligated to the restriction fragments of the DNA. The restriction fragments are then amplified using PCR by using the adapter and restriction sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotide flanking the restriction sites. These amplified fragments are then visualized on a denaturing polyacrylamide gel.

[0224]AFLP analysis has been performed on Salix (Beismann et al., Mol. Ecol. 6:989-993 (1997), the entirety of which is herein incorporated by reference), Acinetobacter (Janssen et al., Int. J. Syst. Bacteriol. 47:1179-1187 (1997), the entirety of which is herein incorporated by reference), Aeromonas popoffi (Huys et al., Int. J. Syst. Bacteriol. 47:1165-1171 (1997), the entirety of which is herein incorporated by reference), rice (McCouch et al., Plant Mol. Biol. 35:89-99 (1997), the entirety of which is herein incorporated by reference; Nandi et al., Mol. Gen. Genet. 255:1-8 (1997), the entirety of which is herein incorporated by reference; Cho et al., Genome 39:373-378 (1996), the entirety of which is herein incorporated by reference), barley (Hordeum vulgare)(Simons et al., Genomics 44:61-70 (1997), the entirety of which is herein incorporated by reference; Waugh et al., Mol. Gen. Genet. 255:311-321 (1997), the entirety of which is herein incorporated by reference; Qi et al., Mol. Gen Genet. 254:330-336 (1997), the entirety of which is herein incorporated by reference; Becker et al., Mol. Gen. Genet. 249:65-73 (1995), the entirety of which is herein incorporated by reference), potato (Van der Voort et al., Mol. Gen. Genet. 255:438-447 (1997), the entirety of which is herein incorporated by reference; Meksem et al., Mol. Gen. Genet. 249:74-81 (1995), the entirety of which is herein incorporated by reference), Phytophthora infestans (Van der Lee et al., Fungal Genet. Biol. 21:278-291 (1997), the entirety of which is herein incorporated by reference), Bacillus anthracis (Keim et al., J. Bacteriol. 179:818-824 (1997), the entirety of which is herein incorporated by reference), Astragalus cremnophylax (Travis et al., Mol. Ecol. 5:735-745 (1996), the entirety of which is herein incorporated by reference), Arabidopsis (Cnops et al., Mol. Gen. Genet. 253:32-41 (1996), the entirety of which is herein incorporated by reference), Escherichia coli (Lin et al., Nucleic Acids Res. 24:3649-3650 (1996), the entirety of which is herein incorporated by reference), Aeromonas (Huys et al., Int. J. Syst. Bacteriol. 46:572-580 (1996), the entirety of which is herein incorporated by reference), nematode (Folkertsma et al., Mol. Plant Microbe Interact. 9:47-54 (1996), the entirety of which is herein incorporated by reference), tomato (Thomas et al., Plant J. 8:785-794 (1995), the entirety of which is herein incorporated by reference) and human (Latorra et al., PCR Methods Appl. 3:351-358 (1994), the entirety of which is herein incorporated by reference). AFLP analysis has also been used for fingerprinting mRNA (Money et al., Nucleic Acids Res. 24:2616-2617 (1996), the entirety of which is herein incorporated by reference; Bachem et al., Plant J. 9:745-753 (1996), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by AFLP analysis or for fingerprinting RNA.

[0225]Polymorphisms may also be found using random amplified polymorphic DNA (RAPD) (Williams et al., Nucl. Acids Res. 18:6531-6535 (1990), the entirety of which is herein incorporated by reference) and cleavable amplified polymorphic sequences (CAPS) (Lyamichev et al., Science 260:778-783 (1993), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention, may be utilized as markers or probes to detect polymorphisms by RAPD or CAPS analysis.

[0226]Through genetic mapping, a fine scale linkage map can be developed using DNA markers and, then, a genomic DNA library of large-sized fragments can be screened with molecular markers linked to the desired trait. Molecular markers are advantageous for agronomic traits that are otherwise difficult to tag, such as resistance to pathogens, insects and nematodes, tolerance to abiotic stress, quality parameters and quantitative traits such as high yield potential.

[0227]The essential requirements for marker-assisted selection in a plant breeding program are: (1) the marker(s) should co-segregate or be closely linked with the desired trait; (2) an efficient means of screening large populations for the molecular marker(s) should be available; and (3) the screening technique should have high reproducibility across laboratories and preferably be economical to use and be user-friendly.

[0228]The genetic linkage of marker molecules can be established by a gene mapping model such as, without limitation, the flanking marker model reported by Lander and Botstein, Genetics 121:185-199 (1989) and the interval mapping, based on maximum likelihood methods described by Lander and Botstein, Genetics 121:185-199 (1989) and implemented in the software package MAPMAKER/QTL (Lincoln and Lander, Mapping Genes Controlling Quantitative Traits Using MAPMAKER/QTL, Whitehead Institute for Biomedical Research, Massachusetts, (1990). Additional software includes Qgene, Version 2.23 (1996), Department of Plant Breeding and Biometry, 266 Emerson Hall, Cornell University, Ithaca, N.Y., the manual of which is herein incorporated by reference in its entirety). Use of Qgene software is a particularly preferred approach.

[0229]A maximum likelihood estimate (MLE) for the presence of a marker is calculated, together with an MLE assuming no QTL effect, to avoid false positives. A log₁₀ of an odds ratio (LOD) is then calculated as: LOD=log₁₀(MLE for the presence of a QTL/MLE given no linked QTL).

[0230]The LOD score essentially indicates how much more likely the data are to have arisen assuming the presence of a QTL than in its absence. The LOD threshold value for avoiding a false positive with a given confidence, say 95%, depends on the number of markers and the length of the genome. Graphs indicating LOD thresholds are set forth in Lander and Botstein, Genetics 121:185-199 (1989) the entirety of which is herein incorporated by reference and further described by Ar s and Moreno-Gonzalez, Plant Breeding, Hayward et al., (eds.) Chapman & Hall, London, pp. 314-331 (1993), the entirety of which is herein incorporated by reference.

[0231]Additional models can be used. Many modifications and alternative approaches to interval mapping have been reported, including the use non-parametric methods (Kruglyak and Lander, Genetics 139:1421-1428 (1995), the entirety of which is herein incorporated by reference). Multiple regression methods or models can be also be used, in which the trait is regressed on a large number of markers (Jansen, Biometrics in Plant Breeding, van Oijen and Jansen (eds.), Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp. 116-124 (1994); Weber and Wricke, Advances in Plant Breeding, Blackwell, Berlin, 16 (1994), both of which is herein incorporated by reference in their entirety). Procedures combining interval mapping with regression analysis, whereby the phenotype is regressed onto a single putative QTL at a given marker interval and at the same time onto a number of markers that serve as `cofactors,` have been reported by Jansen and Stam, Genetics 136:1447-1455 (1994), the entirety of which is herein incorporated by reference and Zeng, Genetics 136:1457-1468 (1994) the entirety of which is herein incorporated by reference. Generally, the use of cofactors reduces the bias and sampling error of the estimated QTL positions (Utz and Melchinger, Biometrics in Plant Breeding, van Oijen and Jansen (eds.) Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp. 195-204 (1994), the entirety of which is herein incorporated by reference, thereby improving the precision and efficiency of QTL mapping (Zeng, Genetics 136:1457-1468 (1994)). These models can be extended to multi-environment experiments to analyze genotype-environment interactions (Jansen et al., Theo. Appl. Genet. 91:33-37 (1995), the entirety of which is herein incorporated by reference).

[0232]Selection of an appropriate mapping populations is important to map construction. The choice of appropriate mapping population depends on the type of marker systems employed (Tanksley et al., Molecular Mapping Plant Chromosomes. Chromosome Structure and Function: Impact of New Concepts, Gustafson and Appels (eds.), Plenum Press, New York, pp 157-173 (1988), the entirety of which is herein incorporated by reference). Consideration must be given to the source of parents (adapted vs. exotic) used in the mapping population. Chromosome pairing and recombination rates can be severely disturbed (suppressed) in wide crosses (adapted x exotic) and generally yield greatly reduced linkage distances. Wide crosses will usually provide segregating populations with a relatively large array of polymorphisms when compared to progeny in a narrow cross (adapted x adapted).

[0233]An F₂ population is the first generation of selfing after the hybrid seed is produced. Usually a single F₁ plant is selfed to generate a population segregating for all the genes in Mendelian (1:2:1) fashion. Maximum genetic information is obtained from a completely classified F₂ population using a codominant marker system (Mather, Measurement of Linkage in Heredity, Methuen and Co., (1938), the entirety of which is herein incorporated by reference). In the case of dominant markers, progeny tests (e.g. F₃, BCF₂) are required to identify the heterozygotes, thus making it equivalent to a completely classified F₂ population. However, this procedure is often prohibitive because of the cost and time involved in progeny testing. Progeny testing of F₂ individuals is often used in map construction where phenotypes do not consistently reflect genotype (e.g. disease resistance) or where trait expression is controlled by a QTL. Segregation data from progeny test populations (e.g. F₃ or BCF₂) can be used in map construction. Marker-assisted selection can then be applied to cross progeny based on marker-trait map associations (F₂, F₃), where linkage groups have not been completely disassociated by recombination events (i.e., maximum disequilibrium).

[0234]Recombinant inbred lines (RIL) (genetically related lines; usually >F₅, developed from continuously selfing F₂ lines towards homozygosity) can be used as a mapping population. Information obtained from dominant markers can be maximized by using RIL because all loci are homozygous or nearly so. Under conditions of tight linkage (i.e., about <10% recombination), dominant and co-dominant markers evaluated in RIL populations provide more information per individual than either marker type in backcross populations (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992), the entirety of which is herein incorporated by reference). However, as the distance between markers becomes larger (i.e., loci become more independent), the information in RIL populations decreases dramatically when compared to codominant markers.

[0235]Backcross populations (e.g., generated from a cross between a successful variety (recurrent parent) and another variety (donor parent) carrying a trait not present in the former) can be utilized as a mapping population. A series of backcrosses to the recurrent parent can be made to recover most of its desirable traits. Thus a population is created consisting of individuals nearly like the recurrent parent but each individual carries varying amounts or mosaic of genomic regions from the donor parent. Backcross populations can be useful for mapping dominant markers if all loci in the recurrent parent are homozygous and the donor and recurrent parent have contrasting polymorphic marker alleles (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992)). Information obtained from backcross populations using either codominant or dominant markers is less than that obtained from F₂ populations because one, rather than two, recombinant gametes are sampled per plant. Backcross populations, however, are more informative (at low marker saturation) when compared to RILs as the distance between linked loci increases in RIL populations (i.e. about 15% recombination). Increased recombination can be beneficial for resolution of tight linkages, but may be undesirable in the construction of maps with low marker saturation.

[0236]Near-isogenic lines (NIL) created by many backcrosses to produce an array of individuals that are nearly identical in genetic composition except for the trait or genomic region under interrogation can be used as a mapping population. In mapping with NILs, only a portion of the polymorphic loci are expected to map to a selected region.

[0237]Bulk segregant analysis (BSA) is a method developed for the rapid identification of linkage between markers and traits of interest (Michelmore et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9828-9832 (1991), the entirety of which is herein incorporated by reference). In BSA, two bulked DNA samples are drawn from a segregating population originating from a single cross. These bulks contain individuals that are identical for a particular trait (resistant or susceptible to particular disease) or genomic region but arbitrary at unlinked regions (i.e. heterozygous). Regions unlinked to the target region will not differ between the bulked samples of many individuals in BSA.

[0238]It is understood that one or more of the nucleic acid molecules of the present invention may be used as molecular markers. It is also understood that one or more of the protein molecules of the present invention may be used as molecular markers.

[0239]In accordance with this aspect of the present invention, a sample nucleic acid is obtained from plants cells or tissues. Any source of nucleic acid may be used. Preferably, the nucleic acid is genomic DNA. The nucleic acid is subjected to restriction endonuclease digestion. For example, one or more nucleic acid molecule or fragment thereof of the present invention can be used as a probe in accordance with the above-described polymorphic methods. The polymorphism obtained in this approach can then be cloned to identify the mutation at the coding region which alters the protein's structure or regulatory region of the gene which affects its expression level.

[0240]In an aspect of the present invention, one or more of the nucleic molecules of the present invention are used to determine the level (i.e., the concentration of mRNA in a sample, etc.) in a plant (preferably maize or soybean) or pattern (i.e., the kinetics of expression, rate of decomposition, stability profile, etc.) of the expression of a protein encoded in part or whole by one or more of the nucleic acid molecule of the present invention (collectively, the "Expression Response" of a cell or tissue). As used herein, the Expression Response manifested by a cell or tissue is said to be "altered" if it differs from the Expression Response of cells or tissues of plants not exhibiting the phenotype. To determine whether an Expression Response is altered, the Expression Response manifested by the cell or tissue of the plant exhibiting the phenotype is compared with that of a similar cell or tissue sample of a plant not exhibiting the phenotype. As will be appreciated, it is not necessary to re-determine the Expression Response of the cell or tissue sample of plants not exhibiting the phenotype each time such a comparison is made; rather, the Expression Response of a particular plant may be compared with previously obtained values of normal plants. As used herein, the phenotype of the organism is any of one or more characteristics of an organism (e.g. disease resistance, pest tolerance, environmental tolerance such as tolerance to abiotic stress, male sterility, quality improvement or yield etc.). A change in genotype or phenotype may be transient or permanent. Also as used herein, a tissue sample is any sample that comprises more than one cell. In a preferred aspect, a tissue sample comprises cells that share a common characteristic (e.g. derived from root, seed, flower, leaf, stem or pollen etc.).

[0241]In one aspect of the present invention, an evaluation can be conducted to determine whether a particular mRNA molecule is present. One or more of the nucleic acid molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention are utilized to detect the presence or quantity of the mRNA species. Such molecules are then incubated with cell or tissue extracts of a plant under conditions sufficient to permit nucleic acid hybridization. The detection of double-stranded probe-mRNA hybrid molecules is indicative of the presence of the mRNA; the amount of such hybrid formed is proportional to the amount of mRNA. Thus, such probes may be used to ascertain the level and extent of the mRNA production in a plant's cells or tissues. Such nucleic acid hybridization may be conducted under quantitative conditions (thereby providing a numerical value of the amount of the mRNA present). Alternatively, the assay may be conducted as a qualitative assay that indicates either that the mRNA is present, or that its level exceeds a user set, predefined value.

[0242]A principle of in situ hybridization is that a labeled, single-stranded nucleic acid probe will hybridize to a complementary strand of cellular DNA or RNA and, under the appropriate conditions, these molecules will form a stable hybrid. When nucleic acid hybridization is combined with histological techniques, specific DNA or RNA sequences can be identified within a single cell. An advantage of in situ hybridization over more conventional techniques for the detection of nucleic acids is that it allows an investigator to determine the precise spatial population (Angerer et al., Dev. Biol. 101:477-484 (1984), the entirety of which is herein incorporated by reference; Angerer et al., Dev. Biol. 112:157-166 (1985), the entirety of which is herein incorporated by reference; Dixon et al., EMBO J. 10:1317-1324 (1991), the entirety of which is herein incorporated by reference). In situ hybridization may be used to measure the steady-state level of RNA accumulation. It is a sensitive technique and RNA sequences present in as few as 5-10 copies per cell can be detected (Hardin et al., J. Mol. Biol. 202:417-431 (1989), the entirety of which is herein incorporated by reference). A number of protocols have been devised for in situ hybridization, each with tissue preparation, hybridization and washing conditions (Meyerowitz, Plant Mol. Biol. Rep. 5:242-250 (1987), the entirety of which is herein incorporated by reference; Cox and Goldberg, In: Plant Molecular Biology: A Practical Approach, Shaw (ed.), pp 1-35, IRL Press, Oxford (1988), the entirety of which is herein incorporated by reference; Raikhel et al., In situ RNA hybridization in plant tissues, In: Plant Molecular Biology Manual, vol. B9:1-32, Kluwer Academic Publisher, Dordrecht, Belgium (1989), the entirety of which is herein incorporated by reference).

[0243]In situ hybridization also allows for the localization of proteins within a tissue or cell (Wilkinson, In Situ Hybridization, Oxford University Press, Oxford (1992), the entirety of which is herein incorporated by reference; Langdale, In Situ Hybridization In: The Maize Handbook, Freeling and Walbot (eds.), pp 165-179, Springer-Verlag, New York (1994), the entirety of which is herein incorporated by reference). It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the level or pattern of a transcription factor or mRNA thereof by in situ hybridization.

[0244]Fluorescent in situ hybridization allows the localization of a particular DNA sequence along a chromosome which is useful, among other uses, for gene mapping, following chromosomes in hybrid lines or detecting chromosomes with translocations, transversions or deletions. In situ hybridization has been used to identify chromosomes in several plant species (Griffor et al., Plant Mol. Biol. 17:101-109 (1991), the entirety of which is herein incorporated by reference; Gustafson et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:1899-1902 (1990), herein incorporated by reference; Mukai and Gill, Genome 34:448-452 (1991), the entirety of which is herein incorporated by reference; Schwarzacher and Heslop-Harrison, Genome 34:317-323 (1991); Wang et al., Jpn. J. Genet. 66:313-316 (1991), the entirety of which is herein incorporated by reference; Parra and Windle, Nature Genetics 5:17-21 (1993), the entirety of which is herein incorporated by reference). It is understood that the nucleic acid molecules of the present invention may be used as probes or markers to localize sequences along a chromosome.

[0245]Another method to localize the expression of a molecule is tissue printing. Tissue printing provides a way to screen, at the same time on the same membrane many tissue sections from different plants or different developmental stages. Tissue-printing procedures utilize films designed to immobilize proteins and nucleic acids. In essence, a freshly cut section of a tissue is pressed gently onto nitrocellulose paper, nylon membrane or polyvinylidene difluoride membrane. Such membranes are commercially available (e.g. Millipore, Bedford, Mass. U.S.A.). The contents of the cut cell transfer onto the membrane and the contents and are immobilized to the membrane. The immobilized contents form a latent print that can be visualized with appropriate probes. When a plant tissue print is made on nitrocellulose paper, the cell walls leave a physical print that makes the anatomy visible without further treatment (Varner and Taylor, Plant Physiol. 91:31-33 (1989), the entirety of which is herein incorporated by reference).

[0246]Tissue printing on substrate films is described by Daoust, Exp. Cell Res. 12:203-211 (1957), the entirety of which is herein incorporated by reference, who detected amylase, protease, ribonuclease and deoxyribonuclease in animal tissues using starch, gelatin and agar films. These techniques can be applied to plant tissues (Yomo and Taylor, Planta 112:35-43 (1973); the entirety of which is herein incorporated by reference; Harris and Chrispeels, Plant Physiol. 56:292-299 (1975), the entirety of which is herein incorporated by reference). Advances in membrane technology have increased the range of applications of Daoust's tissue-printing techniques allowing (Cassab and Vamer, J. Cell. Biol. 105:2581-2588 (1987), the entirety of which is herein incorporated by reference) the histochemical localization of various plant enzymes and deoxyribonuclease on nitrocellulose paper and nylon (Spruce et al., Phytochemistry 26:2901-2903 (1987), the entirety of which is herein incorporated by reference; Barres et al., Neuron 5:527-544 (1990), the entirety of which is herein incorporated by reference; Reid and Pont-Lezica, Tissue Printing: Tools for the Study of Anatomy, Histochemistry and Gene Expression, Academic Press, New York, N.Y. (1992), the entirety of which is herein incorporated by reference; Reid et al., Plant Physiol. 93:160-165 (1990), the entirety of which is herein incorporated by reference; Ye et al., Plant J. 1:175-183 (1991), the entirety of which is herein incorporated by reference).

[0247]It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the presence or quantity of a transcription factor by tissue printing.

[0248]Further it is also understood that any of the nucleic acid molecules of the present invention may be used as marker nucleic acids and or probes in connection with methods that require probes or marker nucleic acids. As used herein, a probe is an agent that is utilized to determine an attribute or feature (e.g. presence or absence, location, correlation, etc.) of a molecule, cell, tissue or plant. As used herein, a marker nucleic acid is a nucleic acid molecule that is utilized to determine an attribute or feature (e.g., presence or absence, location, correlation, etc.) or a molecule, cell, tissue or plant.

[0249]A microarray-based method for high-throughput monitoring of plant gene expression may be utilized to measure gene-specific hybridization targets. This `chip`-based approach involves using microarrays of nucleic acid molecules as gene-specific hybridization targets to quantitatively measure expression of the corresponding plant genes (Schena et al., Science 270:467-470 (1995), the entirety of which is herein incorporated by reference; Shalon, Ph.D. Thesis, Stanford University (1996), the entirety of which is herein incorporated by reference). Every nucleotide in a large sequence can be queried at the same time. Hybridization can be used to efficiently analyze nucleotide sequences.

[0250]Several microarray methods have been described. One method compares the sequences to be analyzed by hybridization to a set of oligonucleotides representing all possible subsequences (Bains and Smith, J. Theor. Biol. 135:303-307 (1989), the entirety of which is herein incorporated by reference). A second method hybridizes the sample to an array of oligonucleotide or cDNA molecules. An array consisting of oligonucleotides complementary to subsequences of a target sequence can be used to determine the identity of a target sequence, measure its amount and detect differences between the target and a reference sequence. Nucleic acid molecules microarrays may also be screened with protein molecules or fragments thereof to determine nucleic acid molecules that specifically bind protein molecules or fragments thereof.

[0251]The microarray approach may be used with polypeptide targets (U.S. Pat. No. 5,445,934; U.S. Pat. No. 5,143,854; U.S. Pat. No. 5,079,600; U.S. Pat. No. 4,923,901, all of which are herein incorporated by reference in their entirety). Essentially, polypeptides are synthesized on a substrate (microarray) and these polypeptides can be screened with either protein molecules or fragments thereof or nucleic acid molecules in order to screen for either protein molecules or fragments thereof or nucleic acid molecules that specifically bind the target polypeptides. (Fodor et al., Science 251:767-773 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules or protein or fragments thereof of the present invention may be utilized in a microarray based method.

[0252]In a preferred embodiment of the present invention microarrays may be prepared that comprise nucleic acid molecules where such nucleic acid molecules encode at least one, preferably at least two, more preferably at least three tetrapyrrole pathway enzymes. In a preferred embodiment the nucleic acid molecules are selected from the group consisting of a putative maize or soybean chlorophyll synthetase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or fragment thereof, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or fragment thereof.

[0253]Site directed mutagenesis may be utilized to modify nucleic acid sequences, particularly as it is a technique that allows one or more of the amino acids encoded by a nucleic acid molecule to be altered (e.g. a threonine to be replaced by a methionine). Three basic methods for site directed mutagenesis are often employed. These are cassette mutagenesis (Wells et al., Gene 34:315-323 (1985), the entirety of which is herein incorporated by reference), primer extension (Gilliam et al., Gene 12:129-137 (1980), the entirety of which is herein incorporated by reference; Zoller and Smith, Methods Enzymol. 100:468-500 (1983), the entirety of which is herein incorporated by reference; Dalbadie-McFarland et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:6409-6413 (1982), the entirety of which is herein incorporated by reference) and methods based upon PCR (Scharf et al., Science 233:1076-1078 (1986), the entirety of which is herein incorporated by reference; Higuchi et al., Nucleic Acids Res. 16:7351-7367 (1988), the entirety of which is herein incorporated by reference). Site directed mutagenesis approaches are also described in European Patent 0 385 962, the entirety of which is herein incorporated by reference; European Patent 0 359 472, the entirety of which is herein incorporated by reference; and PCT Patent Application WO 93/07278, the entirety of which is herein incorporated by reference.

[0254]Site directed mutagenesis strategies have been applied to plants for both in vitro as well as in vivo site directed mutagenesis (Lanz et al., J. Biol. Chem. 266:9971-9976 (1991), the entirety of which is herein incorporated by reference; Kovgan and Zhdanov, Biotekhnologiya 5:148-154; No. 207160n, Chemical Abstracts 110:225 (1989), the entirety of which is herein incorporated by reference; Ge et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:4037-4041 (1989), the entirety of which is herein incorporated by reference; Zhu et al., J. Biol. Chem. 271:18494-18498 (1996), the entirety of which is herein incorporated by reference; Chu et al., Biochemistry 33:6150-6157 (1994), the entirety of which is herein incorporated by reference; Small et al., EMBO J. 11:1291-1296 (1992), the entirety of which is herein incorporated by reference; Cho et al., Mol. Biotechnol. 8:13-16 (1997), the entirety of which is herein incorporated by reference; Kita et al., J. Biol. Chem. 271:26529-26535 (1996), the entirety of which is herein incorporated by reference, Jin et al., Mol. Microbiol. 7:555-562 (1993), the entirety of which is herein incorporated by reference; Hatfield and Vierstra, J. Biol. Chem. 267:14799-14803 (1992), the entirety of which is herein incorporated by reference; Zhao et al., Biochemistry 31:5093-5099 (1992), the entirety of which is herein incorporated by reference).

[0255]Any of the nucleic acid molecules of the present invention may either be modified by site directed mutagenesis or used as, for example, nucleic acid molecules that are used to target other nucleic acid molecules for modification. It is understood that mutants with more than one altered nucleotide can be constructed using techniques that practitioners are familiar with such as isolating restriction fragments and ligating such fragments into an expression vector (see, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989)).

[0256]Sequence-specific DNA-binding proteins play a role in the regulation of transcription. The isolation of recombinant cDNAs encoding these proteins facilitates the biochemical analysis of their structural and functional properties. Genes encoding such DNA-binding proteins have been isolated using classical genetics (Vollbrecht et al., Nature 350: 241-243 (1991), the entirety of which is herein incorporated by reference) and molecular biochemical approaches, including the screening of recombinant cDNA libraries with antibodies (Landschulz et al., Genes Dev. 2:786-800 (1988), the entirety of which is herein incorporated by reference) or DNA probes (Bodner et al., Cell 55:505-518 (1988), the entirety of which is herein incorporated by reference). In addition, an in situ screening procedure has been used and has facilitated the isolation of sequence-specific DNA-binding proteins from various plant species (Gilmartin et al., Plant Cell 4:839-849 (1992), the entirety of which is herein incorporated by reference; Schindler et al., EMBO J. 11:1261-1273 (1992), the entirety of which is herein incorporated by reference). An in situ screening protocol does not require the purification of the protein of interest (Vinson et al., Genes Dev. 2:801-806 (1988), the entirety of which is herein incorporated by reference; Singh et al., Cell 52:415-423 (1988), the entirety of which is herein incorporated by reference).

[0257]Two steps may be employed to characterize DNA-protein interactions. The first is to identify promoter fragments that interact with DNA-binding proteins, to titrate binding activity, to determine the specificity of binding and to determine whether a given DNA-binding activity can interact with related DNA sequences (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2^nd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)). Electrophoretic mobility-shift assay is a widely used assay. The assay provides a rapid and sensitive method for detecting DNA-binding proteins based on the observation that the mobility of a DNA fragment through a nondenaturing, low-ionic strength polyacrylamide gel is retarded upon association with a DNA-binding protein (Fried and Crother, Nucleic Acids Res. 9:6505-6525 (1981), the entirety of which is herein incorporated by reference). When one or more specific binding activities have been identified, the exact sequence of the DNA bound by the protein may be determined. Several procedures for characterizing protein/DNA-binding sites are used, including methylation and ethylation interference assays (Maxam and Gilbert, Methods Enzymol. 65:499-560 (1980), the entirety of which is herein incorporated by reference; Wissman and Hillen, Methods Enzymol. 208:365-379 (1991), the entirety of which is herein incorporated by reference), footprinting techniques employing DNase I (Galas and Schmitz, Nucleic Acids Res. 5:3157-3170 (1978), the entirety of which is herein incorporated by reference), 1,10-phenanthroline-copper ion methods (Sigman et al., Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference) and hydroxyl radicals methods (Dixon et al., Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention may be utilized to identify a protein or fragment thereof that specifically binds to a nucleic acid molecule of the present invention. It is also understood that one or more of the protein molecules or fragments thereof of the present invention may be utilized to identify a nucleic acid molecule that specifically binds to it.

[0258]A two-hybrid system is based on the fact that many cellular functions are carried out by proteins, such as transcription factors, that interact (physically) with one another. Two-hybrid systems have been used to probe the function of new proteins (Chien et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9578-9582 (1991) the entirety of which is herein incorporated by reference; Durfee et al., Genes Dev. 7:555-569 (1993) the entirety of which is herein incorporated by reference; Choi et al., Cell 78:499-512 (1994), the entirety of which is herein incorporated by reference; Kranz et al., Genes Dev. 8:313-327 (1994), the entirety of which is herein incorporated by reference).

[0259]Interaction mating techniques have facilitated a number of two-hybrid studies of protein-protein interaction. Interaction mating has been used to examine interactions between small sets of tens of proteins (Finley and Brent, Proc. Natl. Acad. Sci. (U.S.A.) 91:12098-12984 (1994), the entirety of which is herein incorporated by reference), larger sets of hundreds of proteins (Bendixen et al., Nucl. Acids Res. 22:1778-1779 (1994), the entirety of which is herein incorporated by reference) and to comprehensively map proteins encoded by a small genome (Bartel et al., Nature Genetics 12:72-77 (1996), the entirety of which is herein incorporated by reference). This technique utilizes proteins fused to the DNA-binding domain and proteins fused to the activation domain. They are expressed in two different haploid yeast strains of opposite mating type and the strains are mated to determine if the two proteins interact. Mating occurs when haploid yeast strains come into contact and result in the fusion of the two haploids into a diploid yeast strain. An interaction can be determined by the activation of a two-hybrid reporter gene in the diploid strain. An advantage of this technique is that it reduces the number of yeast transformations needed to test individual interactions. It is understood that the protein-protein interactions of protein or fragments thereof of the present invention may be investigated using the two-hybrid system and that any of the nucleic acid molecules of the present invention that encode such proteins or fragments thereof may be used to transform yeast in the two-hybrid system.

[0260](a) Plant Constructs and Plant Transformants

[0261]One or more of the nucleic acid molecules of the present invention may be used in plant transformation or transfection. Exogenous genetic material may be transferred into a plant cell and the plant cell regenerated into a whole, fertile or sterile plant. Exogenous genetic material is any genetic material, whether naturally occurring or otherwise, from any source that is capable of being inserted into any organism. Such genetic material may be transferred into either monocotyledons and dicotyledons including, but not limited to maize (pp 63-69), soybean (pp 50-60), Arabidopsis (p 45), phaseolus (pp 47-49), peanut (pp 49-50), alfalfa (p 60), wheat (pp 69-71), rice (pp 72-79), oat (pp 80-81), sorghum (p 83), rye (p 84), tritordeum (p 84), millet (p 85), fescue (p 85), perennial ryegrass (p 86), sugarcane (p 87), cranberry (p 101), papaya (pp 101-102), banana (p 103), banana (p 103), muskmelon (p 104), apple (p 104), cucumber (p 105), dendrobium (p 109), gladiolus (p 110), chrysanthemum (p 110), liliacea (p 111), cotton (pp 113-114), eucalyptus (p 115), sunflower (p 118), canola (p 118), turfgrass (p 121), sugarbeet (p 122), coffee (p 122) and dioscorea (p 122), (Christou, In: Particle Bombardment for Genetic Engineering of Plants, Biotechnology Intelligence Unit. Academic Press, San Diego, Calif. (1996), the entirety of which is herein incorporated by reference).

[0262]Transfer of a nucleic acid that encodes for a protein can result in overexpression of that protein in a transformed cell or transgenic plant. One or more of the proteins or fragments thereof encoded by nucleic acid molecules of the present invention may be overexpressed in a transformed cell or transformed plant. Particularly, any of the transcription factors or fragments thereof may be overexpressed in a transformed cell or transgenic plant. Such overexpression may be the result of transient or stable transfer of the exogenous genetic material.

[0263]Exogenous genetic material may be transferred into a plant cell and the plant cell by the use of a DNA vector or construct designed for such a purpose. Design of such a vector is generally within the skill of the art (See, Plant Molecular Biology: A Laboratory Manual, Clark (ed.), Springier, New York (1997), the entirety of which is herein incorporated by reference).

[0264]A construct or vector may include a plant promoter to express the protein or protein fragment of choice. A number of promoters which are active in plant cells have been described in the literature. These include the nopaline synthase (NOS) promoter (Ebert et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:5745-5749 (1987), the entirety of which is herein incorporated by reference), the octopine synthase (OCS) promoter (which are carried on tumor-inducing plasmids of Agrobacterium tumefaciens), the caulimovirus promoters such as the cauliflower mosaic virus (CaMV) 19S promoter (Lawton et al., Plant Mol. Biol. 9:315-324 (1987), the entirety of which is herein incorporated by reference) and the CAMV 35S promoter (Odell et al., Nature 313:810-812 (1985), the entirety of which is herein incorporated by reference), the figwort mosaic virus 35S-promoter, the light-inducible promoter from the small subunit of ribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the Adh promoter (Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:6624-6628 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase promoter (Yang et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:4144-4148 (1990), the entirety of which is herein incorporated by reference), the R gene complex promoter (Chandler et al., The Plant Cell 1:1175-1183 (1989), the entirety of which is herein incorporated by reference) and the chlorophyll a/b binding protein gene promoter, etc. These promoters have been used to create DNA constructs which have been expressed in plants; see, e.g., PCT publication WO 84/02913, herein incorporated by reference in its entirety.

[0265]Promoters which are known or are found to cause transcription of DNA in plant cells can be used in the present invention. Such promoters may be obtained from a variety of sources such as plants and plant viruses. It is preferred that the particular promoter selected should be capable of causing sufficient expression to result in the production of an effective amount of the transcription factor to cause the desired phenotype. In addition to promoters that are known to cause transcription of DNA in plant cells, other promoters may be identified for use in the current invention by screening a plant cDNA library for genes which are selectively or preferably expressed in the target tissues or cells.

[0266]For the purpose of expression in source tissues of the plant, such as the leaf, seed, root or stem, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. For this purpose, one may choose from a number of promoters for genes with tissue- or cell-specific or -enhanced expression. Examples of such promoters reported in the literature include the chloroplast glutamine synthetase GS2 promoter from pea (Edwards et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:3459-3463 (1990), herein incorporated by reference in its entirety), the chloroplast fructose-1,6-biphosphatase (FBPase) promoter from wheat (Lloyd et al., Mol. Gen. Genet. 225:209-216 (1991), herein incorporated by reference in its entirety), the nuclear photosynthetic ST-LS1 promoter from potato (Stockhaus et al., EMBO J. 8:2445-2451 (1989), herein incorporated by reference in its entirety), the serine/threonine kinase (PAL) promoter and the glucoamylase (CHS) promoter from Arabidopsis thaliana. Also reported to be active in photosynthetically active tissues are the ribulose-1,5-bisphosphate carboxylase (RbcS) promoter from eastern larch (Larix laricina), the promoter for the cab gene, cab6, from pine (Yamamoto et al., Plant Cell Physiol. 35:773-778 (1994), herein incorporated by reference in its entirety), the promoter for the Cab-1 gene from wheat (Fejes et al., Plant Mol. Biol. 15:921-932 (1990), herein incorporated by reference in its entirety), the promoter for the CAB-1 gene from spinach (Lubberstedt et al., Plant Physiol. 104:997-1006 (1994), herein incorporated by reference in its entirety), the promoter for the cab1R gene from rice (Luan et al., Plant Cell. 4:971-981 (1992), the entirety of which is herein incorporated by reference), the pyruvate, orthophosphate dikinase (PPDK) promoter from maize (Matsuoka et al., Proc. Natl. Acad. Sci. (U.S.A.) 90: 9586-9590 (1993), herein incorporated by reference in its entirety), the promoter for the tobacco Lhcb 1*2 gene (Cerdan et al., Plant Mol. Biol. 33:245-255 (1997), herein incorporated by reference in its entirety), the Arabidopsis thaliana SUC2 sucrose-H+ symporter promoter (Truernit et al., Planta. 196:564-570 (1995), herein incorporated by reference in its entirety) and the promoter for the thylakoid membrane proteins from spinach (psaD, psaF, psaE, PC, FNR, atpC, atpD, cab, rbcS). Other promoters for the chlorophyll a/b-binding proteins may also be utilized in the present invention, such as the promoters for LhcB gene and PsbP gene from white mustard (Sinapis alba; Kretsch et al., Plant Mol. Biol. 28:219-229 (1995), the entirety of which is herein incorporated by reference).

[0267]For the purpose of expression in sink tissues of the plant, such as the tuber of the potato plant, the fruit of tomato, or the seed of maize, wheat, rice and barley, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. A number of promoters for genes with tuber-specific or -enhanced expression are known, including the class I patatin promoter (Bevan et al., EMBO J. 8:1899-1906 (1986); Jefferson et al., Plant Mol. Biol. 14:995-1006 (1990), both of which are herein incorporated by reference in its entirety), the promoter for the potato tuber ADPGPP genes, both the large and small subunits, the sucrose synthase promoter (Salanoubat and Belliard, Gene. 60:47-56 (1987), Salanoubat and Belliard, Gene. 84:181-185 (1989), both of which are incorporated by reference in their entirety), the promoter for the major tuber proteins including the 22 kd protein complexes and proteinase inhibitors (Hannapel, Plant Physiol. 101:703-704 (1993), herein incorporated by reference in its entirety), the promoter for the granule bound starch synthase gene (GBSS) (Visser et al., Plant Mol. Biol. 17:691-699 (1991), herein incorporated by reference in its entirety) and other class I and II patatins promoters (Koster-Topfer et al., Mol Gen Genet. 219:390-396 (1989); Mignery et al., Gene. 62:27-44 (1988), both of which are herein incorporated by reference in their entirety).

[0268]Other promoters can also be used to express a transcription factor or fragment thereof in specific tissues, such as seeds or fruits. The promoter for β-conglycinin (Chen et al., Dev. Genet. 10: 112-122 (1989), herein incorporated by reference in its entirety) or other seed-specific promoters such as the napin and phaseolin promoters, can be used. The zeins are a group of storage proteins found in maize endosperm. Genomic clones for zein genes have been isolated (Pedersen et al., Cell 29:1015-1026 (1982), herein incorporated by reference in its entirety) and the promoters from these clones, including the 15 kD, 16 kD, 19 kD, 22 kD, 27 kD and γ genes, could also be used. Other promoters known to function, for example, in maize include the promoters for the following genes: waxy, Brittle, Shrunken 2, Branching enzymes I and II, starch synthases, debranching enzymes, oleosins, glutelins and sucrose synthases. A particularly preferred promoter for maize endosperm expression is the promoter for the glutelin gene from rice, more particularly the Osgt-1 promoter (Zheng et al., Mol. Cell Biol. 13:5829-5842 (1993), herein incorporated by reference in its entirety). Examples of promoters suitable for expression in wheat include those promoters for the ADPglucose pyrosynthase (ADPGPP) subunits, the granule bound and other starch synthase, the branching and debranching enzymes, the embryogenesis-abundant proteins, the gliadins and the glutenins. Examples of such promoters in rice include those promoters for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases and the glutelins. A particularly preferred promoter is the promoter for rice glutelin, Osgt-1. Examples of such promoters for barley include those for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases, the hordeins, the embryo globulins and the aleurone specific proteins.

[0269]Root specific promoters may also be used. An example of such a promoter is the promoter for the acid chitinase gene (Samac et al., Plant Mol. Biol. 25:587-596 (1994), the entirety of which is herein incorporated by reference). Expression in root tissue could also be accomplished by utilizing the root specific subdomains of the CaMV35S promoter that have been identified (Lam et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:7890-7894 (1989), herein incorporated by reference in its entirety). Other root cell specific promoters include those reported by Conkling et al. (Conkling et al., Plant Physiol. 93:1203-1211 (1990), the entirety of which is herein incorporated by reference).

[0270]Additional promoters that may be utilized are described, for example, in U.S. Pat. Nos. 5,378,619; 5,391,725; 5,428,147; 5,447,858; 5,608,144; 5,608,144; 5,614,399; 5,633,441; 5,633,435; and 4,633,436, all of which are herein incorporated in their entirety. In addition, a tissue specific enhancer may be used (Fromm et al., The Plant Cell 1:977-984 (1989), the entirety of which is herein incorporated by reference).

[0271]Constructs or vectors may also include with the coding region of interest a nucleic acid sequence that acts, in whole or in part, to terminate transcription of that region. For example, such sequences have been isolated including the Tr7 3' sequence and the NOS 3' sequence (Ingelbrecht et al., The Plant Cell 1:671-680 (1989), the entirety of which is herein incorporated by reference; Bevan et al., Nucleic Acids Res. 11:369-385 (1983), the entirety of which is herein incorporated by reference), or the like.

[0272]A vector or construct may also include regulatory elements. Examples of such include the Adh intron 1 (Callis et al., Genes and Develop. 1:1183-1200 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase intron (Vasil et al., Plant Physiol. 91:1575-1579 (1989), the entirety of which is herein incorporated by reference) and the TMV omega element (Gallie et al., The Plant Cell 1:301-311 (1989), the entirety of which is herein incorporated by reference). These and other regulatory elements may be included when appropriate.

[0273]A vector or construct may also include a selectable marker. Selectable markers may also be used to select for plants or plant cells that contain the exogenous genetic material. Examples of such include, but are not limited to, a neo gene (Potrykus et al., Mol. Gen. Genet. 199:183-188 (1985), the entirety of which is herein incorporated by reference) which codes for kanamycin resistance and can be selected for using kanamycin, G418, etc.; a bar gene which codes for bialaphos resistance; a mutant EPSP synthase gene (Hinchee et al., Bio/Technology 6:915-922 (1988), the entirety of which is herein incorporated by reference) which encodes glyphosate resistance; a nitrilase gene which confers resistance to bromoxynil (Stalker et al., J. Biol. Chem. 263:6310-6314 (1988), the entirety of which is herein incorporated by reference); a mutant acetolactate synthase gene (ALS) which confers imidazolinone or sulphonylurea resistance (European Patent Application 154,204 (Sep. 11, 1985), the entirety of which is herein incorporated by reference); and a methotrexate resistant DHFR gene (Thillet et al., J. Biol. Chem. 263:12500-12508 (1988), the entirety of which is herein incorporated by reference).

[0274]A vector or construct may also include a transit peptide. Incorporation of a suitable chloroplast transit peptide may also be employed (European Patent Application Publication Number 0218571, the entirety of which is herein incorporated by reference). Translational enhancers may also be incorporated as part of the vector DNA. DNA constructs could contain one or more 5' non-translated leader sequences which may serve to enhance expression of the gene products from the resulting mRNA transcripts. Such sequences may be derived from the promoter selected to express the gene or can be specifically modified to increase translation of the mRNA. Such regions may also be obtained from viral RNAs, from suitable eukaryotic genes, or from a synthetic gene sequence. For a review of optimizing expression of transgenes, see Koziel et al., Plant Mol. Biol. 32:393-405 (1996), the entirety of which is herein incorporated by reference.

[0275]A vector or construct may also include a screenable marker. Screenable markers may be used to monitor expression. Exemplary screenable markers include a β-glucuronidase or uidA gene (GUS) which encodes an enzyme for which various chromogenic substrates are known (Jefferson, Plant Mol. Biol, Rep. 5:387-405 (1987), the entirety of which is herein incorporated by reference; Jefferson et al., EMBO J. 6:3901-3907 (1987), the entirety of which is herein incorporated by reference); an R-locus gene, which encodes a product that regulates the production of anthocyanin pigments (red color) in plant tissues (Dellaporta et al., Stadler Symposium 11:263-282 (1988), the entirety of which is herein incorporated by reference); a β-lactamase gene (Sutcliffe et al., Proc. Natl. Acad. Sci. (U.S.A.) 75:3737-3741 (1978), the entirety of which is herein incorporated by reference), a gene which encodes an enzyme for which various chromogenic substrates are known (e.g., PADAC, a chromogenic cephalosporin); a luciferase gene (Ow et al., Science 234:856-859 (1986), the entirety of which is herein incorporated by reference); a xylE gene (Zukowsky et al., Proc. Natl. Acad. Sci. (U.S.A.) 80:1101-1105 (1983), the entirety of which is herein incorporated by reference) which encodes a catechol dioxygenase that can convert chromogenic catechols; an α-amylase gene (Ikatu et al., Bio/Technol. 8:241-242 (1990), the entirety of which is herein incorporated by reference); a tyrosinase gene (Katz et al., J. Gen. Microbiol. 129:2703-2714 (1983), the entirety of which is herein incorporated by reference) which encodes an enzyme capable of oxidizing tyrosine to DOPA and dopaquinone which in turn condenses to melanin; an α-galactosidase, which will turn a chromogenic α-galactose substrate.

[0276]Included within the terms "selectable or screenable marker genes" are also genes which encode a secretable marker whose secretion can be detected as a means of identifying or selecting for transformed cells. Examples include markers which encode a secretable antigen that can be identified by antibody interaction, or even secretable enzymes which can be detected catalytically. Secretable proteins fall into a number of classes, including small, diffusible proteins which are detectable, (e.g., by ELISA), small active enzymes which are detectable in extracellular solution (e.g., α-amylase, β-lactamase, phosphinothricin transferase), or proteins which are inserted or trapped in the cell wall (such as proteins which include a leader sequence such as that found in the expression unit of extension or tobacco PR-S). Other possible selectable and/or screenable marker genes will be apparent to those of skill in the art.

[0277]There are many methods for introducing transforming nucleic acid molecules into plant cells. Suitable methods are believed to include virtually any method by which nucleic acid molecules may be introduced into a cell, such as by Agrobacterium infection or direct delivery of nucleic acid molecules such as, for example, by PEG-mediated transformation, by electroporation or by acceleration of DNA coated particles, etc (Potrykus, Ann. Rev. Plant Physiol. Plant Mol. Biol. 42:205-225 (1991), the entirety of which is herein incorporated by reference; Vasil, Plant Mol. Biol. 25:925-937 (1994), the entirety of which is herein incorporated by reference). For example, electroporation has been used to transform maize protoplasts (Fromm et al., Nature 312:791-793 (1986), the entirety of which is herein incorporated by reference).

[0278]Other vector systems suitable for introducing transforming DNA into a host plant cell include but are not limited to binary artificial chromosome (BIBAC) vectors (Hamilton et al., Gene 200:107-116 (1997), the entirety of which is herein incorporated by reference); and transfection with RNA viral vectors (Della-Cioppa et al., Ann. N.Y. Acad. Sci. (1996), 792 (Engineering Plants for Commercial Products and Applications), 57-61, the entirety of which is herein incorporated by reference). Additional vector systems also include plant selectable YAC vectors such as those described in Mullen et al., Molecular Breeding 4:449-457 (1988), the entirety of which is herein incorporated by reference).

[0279]Technology for introduction of DNA into cells is well known to those of skill in the art. Four general methods for delivering a gene into cells have been described: (1) chemical methods (Graham and van der Eb, Virology 54:536-539 (1973), the entirety of which is herein incorporated by reference); (2) physical methods such as microinjection (Capecchi, Cell 22:479-488 (1980), the entirety of which is herein incorporated by reference), electroporation (Wong and Neumann, Biochem. Biophys. Res. Commun. 107:584-587 (1982); Fromm et al., Proc. Natl. Acad. Sci. (U.S.A.) 82:5824-5828 (1985); U.S. Pat. No. 5,384,253, all of which are herein incorporated in their entirety); and the gene gun (Johnston and Tang, Methods Cell Biol. 43:353-365 (1994), the entirety of which is herein incorporated by reference); (3) viral vectors (Clapp, Clin. Perinatol. 20:155-168 (1993); Lu et al., J. Exp. Med. 178:2089-2096 (1993); Eglitis and Anderson, Biotechniques 6:608-614 (1988), all of which are herein incorporated in their entirety); and (4) receptor-mediated mechanisms (Curiel et al., Hum. Gen. Ther. 3:147-154 (1992), Wagner et al., Proc. Natl. Acad. Sci. (USA) 89:6099-6103 (1992), both of which are incorporated by reference in their entirety).

[0280]Acceleration methods that may be used include, for example, microprojectile bombardment and the like. One example of a method for delivering transforming nucleic acid molecules to plant cells is microprojectile bombardment. This method has been reviewed by Yang and Christou (eds.), Particle Bombardment Technology for Gene Transfer, Oxford Press, Oxford, England (1994), the entirety of which is herein incorporated by reference). Non-biological particles (microprojectiles) that may be coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, gold, platinum and the like.

[0281]A particular advantage of microprojectile bombardment, in addition to it being an effective means of reproducibly transforming monocots, is that neither the isolation of protoplasts (Cristou et al., Plant Physiol. 87:671-674 (1988), the entirety of which is herein incorporated by reference) nor the susceptibility of Agrobacterium infection are required. An illustrative embodiment of a method for delivering DNA into maize cells by acceleration is a biolistics α-particle delivery system, which can be used to propel particles coated with DNA through a screen, such as a stainless steel or Nytex screen, onto a filter surface covered with corn cells cultured in suspension. Gordon-Kamm et al., describes the basic procedure for coating tungsten particles with DNA (Gordon-Kamm et al., Plant Cell 2:603-618 (1990), the entirety of which is herein incorporated by reference). The screen disperses the tungsten nucleic acid particles so that they are not delivered to the recipient cells in large aggregates. A particle delivery system suitable for use with the present invention is the helium acceleration PDS-1000/He gun is available from Bio-Rad Laboratories (Bio-Rad, Hercules, Calif.)(Sanford et al., Technique 3:3-16 (1991), the entirety of which is herein incorporated by reference).

[0282]For the bombardment, cells in suspension may be concentrated on filters. Filters containing the cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the gun and the cells to be bombarded.

[0283]Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the acceleration device and the cells to be bombarded. Through the use of techniques set forth herein one may obtain up to 1000 or more foci of cells transiently expressing a marker gene. The number of cells in a focus which express the exogenous gene product 48 hours post-bombardment often range from one to ten and average one to three.

[0284]In bombardment transformation, one may optimize the pre-bombardment culturing conditions and the bombardment parameters to yield the maximum numbers of stable transformants. Both the physical and biological parameters for bombardment are important in this technology. Physical factors are those that involve manipulating the DNA/microprojectile precipitate or those that affect the flight and velocity of either the macro- or microprojectiles. Biological factors include all steps involved in manipulation of cells before and immediately after bombardment, the osmotic adjustment of target cells to help alleviate the trauma associated with bombardment and also the nature of the transforming DNA, such as linearized DNA or intact supercoiled plasmids. It is believed that pre-bombardment manipulations are especially important for successful transformation of immature embryos.

[0285]In another alternative embodiment, plastids can be stably transformed. Methods disclosed for plastid transformation in higher plants include the particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination (Svab et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8526-8530 (1990); Svab and Maliga, Proc. Natl. Acad. Sci. (U.S.A.) 90:913-917 (1993); Staub and Maliga, EMBO J. 12:601-606 (1993); U.S. Pat. Nos. 5,451,513 and 5,545,818, all of which are herein incorporated by reference in their entirety).

[0286]Accordingly, it is contemplated that one may wish to adjust various aspects of the bombardment parameters in small scale studies to fully optimize the conditions. One may particularly wish to adjust physical parameters such as gap distance, flight distance, tissue distance and helium pressure. One may also minimize the trauma reduction factors by modifying conditions which influence the physiological state of the recipient cells and which may therefore influence transformation and integration efficiencies. For example, the osmotic state, tissue hydration and the subculture stage or cell cycle of the recipient cells may be adjusted for optimum transformation. The execution of other routine adjustments will be known to those of skill in the art in light of the present disclosure.

[0287]Agrobacterium-mediated transfer is a widely applicable system for introducing genes into plant cells because the DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art. See, for example the methods described by Fraley et al., Bio/Technology 3:629-635 (1985) and Rogers et al., Methods Enzymol. 153:253-277 (1987), both of which are herein incorporated by reference in their entirety. Further, the integration of the Ti-DNA is a relatively precise process resulting in few rearrangements. The region of DNA to be transferred is defined by the border sequences and intervening DNA is usually inserted into the plant genome as described (Spielmann et al., Mol. Gen. Genet. 205:34 (1986), the entirety of which is herein incorporated by reference).

[0288]Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations as described (Klee et al., In: Plant DNA Infectious Agents, Hohn and Schell (eds.), Springer-Verlag, New York, pp. 179-203 (1985), the entirety of which is herein incorporated by reference. Moreover, technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes and are suitable for present purposes (Rogers et al., Methods Enzymol. 153:253-277 (1987)). In addition, Agrobacterium containing both armed and disarmed Ti genes can be used for the transformations. In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene transfer.

[0289]A transgenic plant formed using Agrobacterium transformation methods typically contains a single gene on one chromosome. Such transgenic plants can be referred to as being heterozygous for the added gene. More preferred is a transgenic plant that is homozygous for the added structural gene; i.e., a transgenic plant that contains two added genes, one gene at the same locus on each chromosome of a chromosome pair. A homozygous transgenic plant can be obtained by sexually mating (selfing) an independent segregant transgenic plant that contains a single added gene, germinating some of the seed produced and analyzing the resulting plants produced for the gene of interest.

[0290]It is also to be understood that two different transgenic plants can also be mated to produce offspring that contain two independently segregating added, exogenous genes. Selfing of appropriate progeny can produce plants that are homozygous for both added, exogenous genes that encode a polypeptide of interest. Back-crossing to a parental plant and out-crossing with a non-transgenic plant are also contemplated, as is vegetative propagation.

[0291]Transformation of plant protoplasts can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation and combinations of these treatments (See, for example, Potrykus et al., Mol. Gen. Genet. 205:193-200 (1986); Lorz et al., Mol. Gen. Genet. 199:178 (1985); Fromm et al., Nature 319:791 (1986); Uchimiya et al., Mol. Gen. Genet. 204:204 (1986); Marcotte et al., Nature 335:454-457 (1988), all of which are herein incorporated by reference in their entirety).

[0292]Application of these systems to different plant strains depends upon the ability to regenerate that particular plant strain from protoplasts. Illustrative methods for the regeneration of cereals from protoplasts are described (Fujimura et al., Plant Tissue Culture Letters 2:74 (1985); Toriyama et al., Theor Appl. Genet. 205:34 (1986); Yamada et al., Plant Cell Rep. 4:85 (1986); Abdullah et al., Biotechnology 4:1087 (1986), all of which are herein incorporated by reference in their entirety).

[0293]To transform plant strains that cannot be successfully regenerated from protoplasts, other ways to introduce DNA into intact cells or tissues can be utilized. For example, regeneration of cereals from immature embryos or explants can be effected as described (Vasil, Biotechnology 6:397 (1988), the entirety of which is herein incorporated by reference). In addition, "particle gun" or high-velocity microprojectile technology can be utilized (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference).

[0294]Using the latter technology, DNA is carried through the cell wall and into the cytoplasm on the surface of small metal particles as described (Klein et al., Nature 328:70 (1987); Klein et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8502-8505 (1988); McCabe et al., Bio/Technology 6:923 (1988), all of which are herein incorporated by reference in their entirety). The metal particles penetrate through several layers of cells and thus allow the transformation of cells within tissue explants.

[0295]Other methods of cell transformation can also be used and include but are not limited to introduction of DNA into plants by direct DNA transfer into pollen (Zhou et al., Methods Enzymol. 101:433 (1983); Hess et al., Intern Rev. Cytol. 107:367 (1987); Luo et al., Plant Mol Biol. Reporter 6:165 (1988), all of which are herein incorporated by reference in their entirety), by direct injection of DNA into reproductive organs of a plant (Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference), or by direct injection of DNA into the cells of immature embryos followed by the rehydration of desiccated embryos (Neuhaus et al., Theor. Appl. Genet. 75:30 (1987), the entirety of which is herein incorporated by reference).

[0296]The regeneration, development and cultivation of plants from single plant protoplast transformants or from various transformed explants is well known in the art (Weissbach and Weissbach, In: Methods for Plant Molecular Biology, Academic Press, San Diego, Calif., (1988), the entirety of which is herein incorporated by reference). This regeneration and growth process typically includes the steps of selection of transformed cells, culturing those individualized cells through the usual stages of embryonic development through the rooted plantlet stage. Transgenic embryos and seeds are similarly regenerated. The resulting transgenic rooted shoots are thereafter planted in an appropriate plant growth medium such as soil.

[0297]The development or regeneration of plants containing the foreign, exogenous gene that encodes a protein of interest is well known in the art. Preferably, the regenerated plants are self-pollinated to provide homozygous transgenic plants. Otherwise, pollen obtained from the regenerated plants is crossed to seed-grown plants of agronomically important lines. Conversely, pollen from plants of these important lines is used to pollinate regenerated plants. A transgenic plant of the present invention containing a desired polypeptide is cultivated using methods well known to one skilled in the art.

[0298]There are a variety of methods for the regeneration of plants from plant tissue. The particular method of regeneration will depend on the starting plant tissue and the particular plant species to be regenerated.

[0299]Methods for transforming dicots, primarily by use of Agrobacterium tumefaciens and obtaining transgenic plants have been published for cotton (U.S. Pat. No. 5,004,863; U.S. Pat. No. 5,159,135; U.S. Pat. No. 5,518,908, all of which are herein incorporated by reference in their entirety); soybean (U.S. Pat. No. 5,569,834; U.S. Pat. No. 5,416,011; McCabe et. al., Biotechnology 6:923 (1988); Christou et al., Plant Physiol. 87:671-674 (1988); all of which are herein incorporated by reference in their entirety); Brassica (U.S. Pat. No. 5,463,174, the entirety of which is herein incorporated by reference); peanut (Cheng et al., Plant Cell Rep. 15:653-657 (1996), McKently et al., Plant Cell Rep. 14:699-703 (1995), all of which are herein incorporated by reference in their entirety); papaya; and pea (Grant et al., Plant Cell Rep. 15:254-258 (1995), the entirety of which is herein incorporated by reference).

[0300]Transformation of monocotyledons using electroporation, particle bombardment and Agrobacterium have also been reported. Transformation and plant regeneration have been achieved in asparagus (Bytebier et al., Proc. Natl. Acad. Sci. (USA) 84:5354 (1987), the entirety of which is herein incorporated by reference); barley (Wan and Lemaux, Plant Physiol 104:37 (1994), the entirety of which is herein incorporated by reference); maize (Rhodes et al., Science 240:204 (1988); Gordon-Kamm et al., Plant Cell 2:603-618 (1990); Fromm et al., Bio/Technology 8:833 (1990); Koziel et al., Bio/Technology 11:194 (1993); Armstrong et al., Crop Science 35:550-557 (1995); all of which are herein incorporated by reference in their entirety); oat (Somers et al., Bio/Technology 10:1589 (1992), the entirety of which is herein incorporated by reference); orchard grass (Horn et al., Plant Cell Rep. 7:469 (1988), the entirety of which is herein incorporated by reference); rice (Toriyama et al., Theor Appl. Genet. 205:34 (1986); Part et al., Plant Mol. Biol. 32:1135-1148 (1996); Abedinia et al., Aust. J. Plant Physiol. 24:133-141 (1997); Zhang and Wu, Theor. Appl. Genet. 76:835 (1988); Zhang et al., Plant Cell Rep. 7:379 (1988); Battraw and Hall, Plant Sci. 86:191-202 (1992); Christou et al., Bio/Technology 9:957 (1991), all of which are herein incorporated by reference in their entirety); rye (De la Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference); sugarcane (Bower and Birch, Plant J. 2:409 (1992), the entirety of which is herein incorporated by reference); tall fescue (Wang et al., Bio/Technology 10:691 (1992), the entirety of which is herein incorporated by reference) and wheat (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference; U.S. Pat. No. 5,631,152, the entirety of which is herein incorporated by reference.)

[0301]Assays for gene expression based on the transient expression of cloned nucleic acid constructs have been developed by introducing the nucleic acid molecules into plant cells by polyethylene glycol treatment, electroporation, or particle bombardment (Marcotte et al., Nature 335:454-457 (1988), the entirety of which is herein incorporated by reference; Marcotte et al., Plant Cell 1:523-532 (1989), the entirety of which is herein incorporated by reference; McCarty et al., Cell 66:895-905 (1991), the entirety of which is herein incorporated by reference; Hattori et al., Genes Dev. 6:609-618 (1992), the entirety of which is herein incorporated by reference; Goff et al., EMBO J. 9:2517-2522 (1990), the entirety of which is herein incorporated by reference). Transient expression systems may be used to functionally dissect gene constructs (see generally, Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995)).

[0302]Any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a permanent or transient manner in combination with other genetic elements such as vectors, promoters, enhancers etc. Further, any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a manner that allows for overexpression of the protein or fragment thereof encoded by the nucleic acid molecule.

[0303]Cosuppression is the reduction in expression levels, usually at the level of RNA, of a particular endogenous gene or gene family by the expression of a homologous sense construct that is capable of transcribing mRNA of the same strandedness as the transcript of the endogenous gene (Napoli et al., Plant Cell 2:279-289 (1990), the entirety of which is herein incorporated by reference; van der Krol et al., Plant Cell 2:291-299 (1990), the entirety of which is herein incorporated by reference). Cosuppression may result from stable transformation with a single copy nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Prolls and Meyer, Plant J. 2:465-475 (1992), the entirety of which is herein incorporated by reference) or with multiple copies of a nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Mittlesten et al., Mol. Gen. Genet. 244:325-330 (1994), the entirety of which is herein incorporated by reference). Genes, even though different, linked to homologous promoters may result in the cosuppression of the linked genes (Vaucheret, C.R. Acad. Sci. III 316:1471-1483 (1993), the entirety of which is herein incorporated by reference).

[0304]This technique has, for example, been applied to generate white flowers from red petunia and tomatoes that do not ripen on the vine. Up to 50% of petunia transformants that contained a sense copy of the glucoamylase (CHS) gene produced white flowers or floral sectors; this was as a result of the post-transcriptional loss of mRNA encoding CHS (Flavell, Proc. Natl. Acad. Sci. (U.S.A.) 91:3490-3496 (1994), the entirety of which is herein incorporated by reference); van Blokland et al., Plant J. 6:861-877 (1994), the entirety of which is herein incorporated by reference). Cosuppression may require the coordinate transcription of the transgene and the endogenous gene and can be reset by a developmental control mechanism (Jorgensen, Trends Biotechnol. 8:340-344 (1990), the entirety of which is herein incorporated by reference; Meins and Kunz, In: Gene Inactivation and Homologous Recombination in Plants, Paszkowski (ed.), pp. 335-348, Kluwer Academic, Netherlands (1994), the entirety of which is herein incorporated by reference).

[0305]It is understood that one or more of the nucleic acids of the present invention may be introduced into a plant cell and transcribed using an appropriate promoter with such transcription resulting in the cosuppression of an endogenous transcription factor.

[0306]Antisense approaches are a way of preventing or reducing gene function by targeting the genetic material (Mol et al., FEBS Lett. 268:427-430 (1990), the entirety of which is herein incorporated by reference). The objective of the antisense approach is to use a sequence complementary to the target gene to block its expression and create a mutant cell line or organism in which the level of a single chosen protein is selectively reduced or abolished. Antisense techniques have several advantages over other `reverse genetic` approaches. The site of inactivation and its developmental effect can be manipulated by the choice of promoter for antisense genes or by the timing of external application or microinjection. Antisense can manipulate its specificity by selecting either unique regions of the target gene or regions where it shares homology to other related genes (Hiatt et al., In: Genetic Engineering, Setlow (ed.), Vol. 11, New York: Plenum 49-63 (1989), the entirety of which is herein incorporated by reference).

[0307]The principle of regulation by antisense RNA is that RNA that is complementary to the target mRNA is introduced into cells, resulting in specific RNA:RNA duplexes being formed by base pairing between the antisense substrate and the target mRNA (Green et al., Annu. Rev. Biochem. 55:569-597 (1986), the entirety of which is herein incorporated by reference). Under one embodiment, the process involves the introduction and expression of an antisense gene sequence. Such a sequence is one in which part or all of the normal gene sequences are placed under a promoter in inverted orientation so that the `wrong` or complementary strand is transcribed into a noncoding antisense RNA that hybridizes with the target mRNA and interferes with its expression (Takayama and Inouye, Crit. Rev. Biochem. Mol. Biol. 25:155-184 (1990), the entirety of which is herein incorporated by reference). An antisense vector is constructed by standard procedures and introduced into cells by transformation, transfection, electroporation, microinjection, infection, etc. The type of transformation and choice of vector will determine whether expression is transient or stable. The promoter used for the antisense gene may influence the level, timing, tissue, specificity, or inducibility of the antisense inhibition.

[0308]It is understood that the activity of a transcription factor in a plant cell may be reduced or depressed by growing a transformed plant cell containing a nucleic acid molecule whose non-transcribed strand encodes a transcription factor or fragment thereof.

[0309]Antibodies have been expressed in plants (Hiatt et al., Nature 342:76-78 (1989), the entirety of which is herein incorporated by reference; Conrad and Fielder, Plant Mol. Biol. 26:1023-1030 (1994), the entirety of which is herein incorporated by reference). Cytoplasmic expression of a scFv (single-chain Fv antibodies) has been reported to delay infection by artichoke mottled crinkle virus. Transgenic plants that express antibodies directed against endogenous proteins may exhibit a physiological effect (Philips et al., EMBO J. 16:4489-4496 (1997), the entirety of which is herein incorporated by reference; Marion-Poll, Trends in Plant Science 2:447-448 (1997), the entirety of which is herein incorporated by reference). For example, expressed anti-abscisic antibodies have been reported to result in a general perturbation of seed development (Philips et al., EMBO J. 16: 4489-4496 (1997)).

[0310]Antibodies that are catalytic may also be expressed in plants (abzymes). The principle behind abzymes is that since antibodies may be raised against many molecules, this recognition ability can be directed toward generating antibodies that bind transition states to force a chemical reaction forward (Persidas, Nature Biotechnology 15:1313-1315 (1997), the entirety of which is herein incorporated by reference; Baca et al., Ann. Rev. Biophys. Biomol. Struct. 26:461-493 (1997), the entirety of which is herein incorporated by reference). The catalytic abilities of abzymes may be enhanced by site directed mutagenesis. Examples of abzymes are, for example, set forth in U.S. Pat. No. 5,658,753; U.S. Pat. No. 5,632,990; U.S. Pat. No. 5,631,137; U.S. Pat. No. 5,602,015; U.S. Pat. No. 5,559,538; U.S. Pat. No. 5,576,174; U.S. Pat. No. 5,500,358; U.S. Pat. No. 5,318,897; U.S. Pat. No. 5,298,409; U.S. Pat. No. 5,258,289 and U.S. Pat. No. 5,194,585, all of which are herein incorporated in their entirety.

[0311]It is understood that any of the antibodies of the present invention may be expressed in plants and that such expression can result in a physiological effect. It is also understood that any of the expressed antibodies may be catalytic.

[0312](b) Fungal Constructs and Fungal Transformants

[0313]The present invention also relates to a fungal recombinant vector comprising exogenous genetic material. The present invention also relates to a fungal cell comprising a fungal recombinant vector. The present invention also relates to methods for obtaining a recombinant fungal host cell comprising introducing into a fungal host cell exogenous genetic material.

[0314]Exogenous genetic material may be transferred into a fungal cell. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragments of either or other nucleic acid molecule of the present invention. The fungal recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the fungal host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the fungal host.

[0315]The fungal vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the fungal cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the fungal host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the fungal host cell and, furthermore, may be non-encoding or encoding sequences.

[0316]For autonomous replication, the vector may further comprise an origin of replication enabling the vector to replicate autonomously in the host cell in question. Examples of origin of replications for use in a yeast host cell are the 2 micron origin of replication and the combination of CEN3 and ARS 1. Any origin of replication may be used which is compatible with the fungal host cell of choice.

[0317]The fungal vectors of the present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. The selectable marker may be selected from the group including, but not limited to, amdS (acetamidase), argB (omithine carbamoyltransferase), bar (phosphinothricin acetyltransferase), hygB (hygromycin phosphotransferase), niaD (nitrate reductase), pyrG (orotidine-5'-phosphate decarboxylase) and sC (sulfate adenyltransferase) and trpC (anthranilate synthase). Preferred for use in an Aspergillus cell are the amdS and pyrG markers of Aspergillus nidulans or Aspergillus oryzae and the bar marker of Streptomyces hygroscopicus. Furthermore, selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, the entirety of which is herein incorporated by reference. A nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the fungal host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof.

[0318]A promoter may be any nucleic acid sequence which shows transcriptional activity in the fungal host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell. Examples of suitable promoters for directing the transcription of a nucleic acid construct of the invention in a filamentous fungal host are promoters obtained from the genes encoding Aspergillus oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, Aspergillus niger neutral alpha-amylase, Aspergillus niger acid stable alpha-amylase, Aspergillus niger or Aspergillus awamori glucoamylase (glaA), Rhizomucor miehei lipase, Aspergillus oryzae alkaline protease, Aspergillus oryzae triose phosphate isomerase, Aspergillus nidulans acetamidase and hybrids thereof. In a yeast host, a useful promoter is the Saccharomyces cerevisiae enolase (eno-1) promoter. Particularly preferred promoters are the TAKA amylase, NA2-tpi (a hybrid of the promoters from the genes encoding Aspergillus niger neutral alpha-amylase and Aspergillus oryzae triose phosphate isomerase) and glaA promoters.

[0319]A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a terminator sequence at its 3' terminus. The terminator sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any terminator which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred terminators are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase, Aspergillus niger alpha-glucosidase and Saccharomyces cerevisiae enolase.

[0320]A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5' terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred leaders are obtained from the genes encoding Aspergillus oryzae TAKA amylase and Aspergillus oryzae triose phosphate isomerase.

[0321]A polyadenylation sequence may also be operably linked to the 3' terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the fungal host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention, but particularly preferred polyadenylation sequences are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase and Aspergillus niger alpha-glucosidase.

[0322]To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed protein or fragment thereof within the cell, it is preferred that expression of the protein or fragment thereof gives rise to a product secreted outside the cell. To this end, a protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the fungal host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5' end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof. Alternatively, the 5' end of the coding sequence may contain a signal peptide coding region which is foreign to that portion of the coding sequence which encodes the secreted protein or fragment thereof. The foreign signal peptide may be required where the coding sequence does not normally contain a signal peptide coding region. Alternatively, the foreign signal peptide may simply replace the natural signal peptide to obtain enhanced secretion of the desired protein or fragment thereof. The foreign signal peptide coding region may be obtained from a glucoamylase or an amylase gene from an Aspergillus species, a lipase or proteinase gene from Rhizomucor miehei, the gene for the alpha-factor from Saccharomyces cerevisiae, or the calf preprochymosin gene. An effective signal peptide for fungal host cells is the Aspergillus oryzae TAKA amylase signal, Aspergillus niger neutral amylase signal, the Rhizomucor miehei aspartic proteinase signal, the Humicola lanuginosus cellulase signal, or the Rhizomucor miehei lipase signal. However, any signal peptide capable of permitting secretion of the protein or fragment thereof in a fungal host of choice may be used in the present invention.

[0323]A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be linked to a propeptide coding region. A propeptide is an amino acid sequence found at the amino terminus of a proprotein or proenzyme. Cleavage of the propeptide from the proprotein yields a mature biochemically active protein. The resulting polypeptide is known as a propolypeptide or proenzyme (or a zymogen in some cases). Propolypeptides are generally inactive and can be converted to mature active polypeptides by catalytic or autocatalytic cleavage of the propeptide from the propolypeptide or proenzyme. The propeptide coding region may be native to the protein or fragment thereof or may be obtained from foreign sources. The foreign propeptide coding region may be obtained from the Saccharomyces cerevisiae alpha-factor gene or Myceliophthora thermophila laccase gene (WO 95/33836, the entirety of which is herein incorporated by reference).

[0324]The procedures used to ligate the elements described above to construct the recombinant expression vector of the present invention are well known to one skilled in the art (see, for example, Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd ed., Cold Spring Harbor, N.Y., (1989)).

[0325]The present invention also relates to recombinant fungal host cells produced by the methods of the present invention which are advantageously used with the recombinant vector of the present invention. The cell is preferably transformed with a vector comprising a nucleic acid sequence of the invention followed by integration of the vector into the host chromosome. The choice of fungal host cells will to a large extent depend upon the gene encoding the protein or fragment thereof and its source. The fungal host cell may, for example, be a yeast cell or a filamentous fungal cell.

[0326]"Yeast" as used herein includes Ascosporogenous yeast (Endomycetales), Basidiosporogenous yeast and yeast belonging to the Fungi Imperfecti (Blastomycetes). The Ascosporogenous yeasts are divided into the families Spermophthoraceae and Saccharomycetaceae. The latter is comprised of four subfamilies, Schizosaccharomycoideae (for example, genus Schizosaccharomyces), Nadsonioideae, Lipomycoideae and Saccharomycoideae (for example, genera Pichia, Kluyveromyces and Saccharomyces). The Basidiosporogenous yeasts include the genera Leucosporidim, Rhodosporidium, Sporidiobolus, Filobasidium and Filobasidiella. Yeast belonging to the Fungi Imperfecti are divided into two families, Sporobolomycetaceae (for example, genera Sorobolomyces and Bullera) and Cryptococcaceae (for example, genus Candida). Since the classification of yeast may change in the future, for the purposes of this invention, yeast shall be defined as described in Biology and Activities of Yeast (Skinner et al., Soc. App. Bacteriol. Symposium Series No. 9, (1980), the entirety of which is herein incorporated by reference). The biology of yeast and manipulation of yeast genetics are well known in the art (see, for example, Biochemistry and Genetics of Yeast, Bacil et al. (ed.), 2nd edition, 1987; The Yeasts, Rose and Harrison (eds.), 2nd ed., (1987); and The Molecular Biology of the Yeast Saccharomyces, Strathern et al. (eds.), (1981), all of which are herein incorporated by reference in their entirety).

[0327]"Fungi" as used herein includes the phyla Ascomycota, Basidiomycota, Chytridiomycota and Zygomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK; the entirety of which is herein incorporated by reference) as well as the Oomycota (as cited in Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK) and all mitosporic fungi (Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). Representative groups of Ascomycota include, for example, Neurospora, Eupenicillium (=Penicillium), Emericella (=Aspergillus), Eurotiun (=Aspergillus) and the true yeasts listed above. Examples of Basidiomycota include mushrooms, rusts and smuts. Representative groups of Chytridiomycota include, for example, Allomyces, Blastocladiella, Coelomomyces and aquatic fungi. Representative groups of Oomycota include, for example, Saprolegniomycetous aquatic fungi (water molds) such as Achlya. Examples of mitosporic fungi include Aspergillus, Penicillium, Candida and Alternaria. Representative groups of Zygomycota include, for example, Rhizopus and Mucor.

[0328]"Filamentous fungi" include all filamentous forms of the subdivision Eumycota and Oomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). The filamentous fungi are characterized by a vegetative mycelium composed of chitin, cellulose, glucan, chitosan, mannan and other complex polysaccharides. Vegetative growth is by hyphal elongation and carbon catabolism is obligately aerobic. In contrast, vegetative growth by yeasts such as Saccharomyces cerevisiae is by budding of a unicellular thallus and carbon catabolism may be fermentative.

[0329]In one embodiment, the fungal host cell is a yeast cell. In a preferred embodiment, the yeast host cell is a cell of the species of Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia and Yarrowia. In a preferred embodiment, the yeast host cell is a Saccharomyces cerevisiae cell, a Saccharomyces carlsbergensis, Saccharomyces diastaticus cell, a Saccharomyces douglasii cell, a Saccharomyces kluyveri cell, a Saccharomyces norbensis cell, or a Saccharomyces oviformis cell. In another preferred embodiment, the yeast host cell is a Kluyveromyces lactis cell. In another preferred embodiment, the yeast host cell is a Yarrowia lipolytica cell.

[0330]In another embodiment, the fungal host cell is a filamentous fungal cell. In a preferred embodiment, the filamentous fungal host cell is a cell of the species of, but not limited to, Acremonium, Aspergillus, Fusarium, Humicola, Myceliophthora, Mucor, Neurospora, Penicillium, Thielavia, Tolypocladium and Trichoderma. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus cell. In another preferred embodiment, the filamentous fungal host cell is an Acremonium cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora cell. In another even preferred embodiment, the filamentous fungal host cell is a Mucor cell. In another preferred embodiment, the filamentous fungal host cell is a Neurospora cell. In another preferred embodiment, the filamentous fungal host cell is a Penicillium cell. In another preferred embodiment, the filamentous fungal host cell is a Thielavia cell. In another preferred embodiment, the filamentous fungal host cell is a Tolypocladiun cell. In another preferred embodiment, the filamentous fungal host cell is a Trichoderma cell. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus oryzae cell, an Aspergillus niger cell, an Aspergillus foetidus cell, or an Aspergillus japonicus cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium oxysporum cell or a Fusarium graminearum cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola insolens cell or a Humicola lanuginosus cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora thermophila cell. In a most preferred embodiment, the filamentous fungal host cell is a Mucor miehei cell. In a most preferred embodiment, the filamentous fungal host cell is a Neurospora crassa cell. In a most preferred embodiment, the filamentous fungal host cell is a Penicillium purpurogenum cell. In another most preferred embodiment, the filamentous fungal host cell is a Thielavia terrestris cell. In another most preferred embodiment, the Trichoderma cell is a Trichoderma reesei cell, a Trichoderna viride cell, a Trichoderma longibrachiatum cell, a Trichoderma harzianum cell, or a Trichoderma koningii cell. In a preferred embodiment, the fungal host cell is selected from an A. nidulans cell, an A. niger cell, an A. oryzae cell and an A. sojae cell. In a further preferred embodiment, the fungal host cell is an A. nidulans cell.

[0331]The recombinant fungal host cells of the present invention may further comprise one or more sequences which encode one or more factors that are advantageous in the expression of the protein or fragment thereof, for example, an activator (e.g., a trans-acting factor), a chaperone and a processing protease. The nucleic acids encoding one or more of these factors are preferably not operably linked to the nucleic acid encoding the protein or fragment thereof. An activator is a protein which activates transcription of a nucleic acid sequence encoding a polypeptide (Kudla et al., EMBO 9:1355-1364(1990); Jarai and Buxton, Current Genetics 26:2238-244(1994); Verdier, Yeast 6:271-297(1990), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding an activator may be obtained from the genes encoding Saccharomyces cerevisiae heme activator protein 1 (hap 1), Saccharomyces cerevisiae galactose metabolizing protein 4 (gal4) and Aspergillus nidulans ammonia regulation protein (areA). For further examples, see Verdier, Yeast 6:271-297 (1990); MacKenzie et al., Journal of Gen. Microbiol. 139:2295-2307 (1993), both of which are herein incorporated by reference in their entirety). A chaperone is a protein which assists another protein in folding properly (Hartl et al., TIBS 19:20-25 (1994); Bergeron et al., TIBS 19:124-128 (1994); Demolder et al., J. Biotechnology 32:179-189 (1994); Craig, Science 260:1902-1903(1993); Gething and Sambrook, Nature 355:33-45 (1992); Puig and Gilbert, J Biol. Chem. 269:7764-7771 (1994); Wang and Tsou, FASEB Journal 7:1515-11157 (1993); Robinson et al., Bio/Technology 1:381-384 (1994), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding a chaperone may be obtained from the genes encoding Aspergillus oryzae protein disulphide isomerase, Saccharomyces cerevisiae calnexin, Saccharomyces cerevisiae BiP/GRP78 and Saccharomyces cerevisiae Hsp70. For further examples, see Gething and Sambrook, Nature 355:33-45 (1992); Hartl et al., TIBS 19:20-25 (1994). A processing protease is a protease that cleaves a propeptide to generate a mature biochemically active polypeptide (Enderlin and Ogrydziak, Yeast 10:67-79 (1994); Fuller et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1434-1438 (1989); Julius et al., Cell 37:1075-1089 (1984); Julius et al., Cell 32:839-852 (1983), all of which are incorporated by reference in their entirety). The nucleic acid sequence encoding a processing protease may be obtained from the genes encoding Aspergillus niger Kex2, Saccharomyces cerevisiae dipeptidylaminopeptidase, Saccharomyces cerevisiae Kex2 and Yarrowia lipolytica dibasic processing endoprotease (xpr6). Any factor that is functional in the fungal host cell of choice may be used in the present invention.

[0332]Fungal cells may be transformed by a process involving protoplast formation, transformation of the protoplasts and regeneration of the cell wall in a manner known per se. Suitable procedures for transformation of Aspergillus host cells are described in EP 238 023 and Yelton et al., Proc. Natl. Acad. Sci. (U.S.A.) 81:1470-1474 (1984), both of which are herein incorporated by reference in their entirety. A suitable method of transforming Fusarium species is described by Malardier et al., Gene 78:147-156 (1989), the entirety of which is herein incorporated by reference. Yeast may be transformed using the procedures described by Becker and Guarente, In: Abelson and Simon, (eds.), Guide to Yeast Genetics and Molecular Biology, Methods Enzymol. Volume 194, pp 182-187, Academic Press, Inc., New York; Ito et al., J. Bacteriology 153:163 (1983); Hinnen et al., Proc. Natl. Acad. Sci. (U.S.A.) 75:1920 (1978), all of which are herein incorporated by reference in their entirety.

[0333]The present invention also relates to methods of producing the protein or fragment thereof comprising culturing the recombinant fungal host cells under conditions conducive for expression of the protein or fragment thereof. The fungal cells of the present invention are cultivated in a nutrient medium suitable for production of the protein or fragment thereof using methods known in the art. For example, the cell may be cultivated by shake flask cultivation, small-scale or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermentors performed in a suitable medium and under conditions allowing the protein or fragment thereof to be expressed and/or isolated. The cultivation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art (see, e.g., Bennett and LaSure (eds.), More Gene Manipulations in Fungi, Academic Press, CA, (1991), the entirety of which is herein incorporated by reference). Suitable media are available from commercial suppliers or may be prepared according to published compositions (e.g., in catalogues of the American Type Culture Collection, Manassas, Va.). If the protein or fragment thereof is secreted into the nutrient medium, a protein or fragment thereof can be recovered directly from the medium. If the protein or fragment thereof is not secreted, it is recovered from cell lysates.

[0334]The expressed protein or fragment thereof may be detected using methods known in the art that are specific for the particular protein or fragment. These detection methods may include the use of specific antibodies, formation of an enzyme product, or disappearance of an enzyme substrate. For example, if the protein or fragment thereof has enzymatic activity, an enzyme assay may be used. Alternatively, if polyclonal or monoclonal antibodies specific to the protein or fragment thereof are available, immunoassays may be employed using the antibodies to the protein or fragment thereof. The techniques of enzyme assay and immunoassay are well known to those skilled in the art.

[0335]The resulting protein or fragment thereof may be recovered by methods known in the arts. For example, the protein or fragment thereof may be recovered from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray-drying, evaporation, or precipitation. The recovered protein or fragment thereof may then be further purified by a variety of chromatographic procedures, e.g., ion exchange chromatography, gel filtration chromatography, affinity chromatography, or the like.

[0336](c) Mammalian Constructs and Transformed Mammalian Cells

[0337]The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian host cell exogenous genetic material. The present invention also relates to a mammalian cell comprising a mammalian recombinant vector. The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

[0338]Mammalian cell lines available as hosts for expression are known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC, Manassas, Va.), such as HeLa cells, Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK) cells and a number of other cell lines. Suitable promoters for mammalian cells are also known in the art and include viral promoters such as that from Simian Virus 40 (SV40) (Fiers et al., Nature 273:113 (1978), the entirety of which is herein incorporated by reference), Rous sarcoma virus (RSV), adenovirus (ADV) and bovine papilloma virus (BPV). Mammalian cells may also require terminator sequences and poly-A addition sequences. Enhancer sequences which increase expression may also be included and sequences which promote amplification of the gene may also be desirable (for example methotrexate resistance genes).

[0339]Vectors suitable for replication in mammalian cells may include viral replicons, or sequences which insure integration of the appropriate sequences encoding HCV epitopes into the host genome. For example, another vector used to express foreign DNA is vaccinia virus. In this case, for example, a nucleic acid molecule encoding a protein or fragment thereof is inserted into the vaccinia genome. Techniques for the insertion of foreign DNA into the vaccinia virus genome are known in the art and may utilize, for example, homologous recombination. Such heterologous DNA is generally inserted into a gene which is non-essential to the virus, for example, the thymidine kinase gene (tk), which also provides a selectable marker. Plasmid vectors that greatly facilitate the construction of recombinant viruses have been described (see, for example, Mackett et al, J Virol. 49:857 (1984); Chakrabarti et al., Mol. Cell. Biol. 5:3403 (1985); Moss, In: Gene Transfer Vectors For Mammalian Cells (Miller and Calos, eds., Cold Spring Harbor Laboratory, N.Y., p. 10, (1987); all of which are herein incorporated by reference in their entirety). Expression of the HCV polypeptide then occurs in cells or animals which are infected with the live recombinant vaccinia virus.

[0340]The sequence to be integrated into the mammalian sequence may be introduced into the primary host by any convenient means, which includes calcium precipitated DNA, spheroplast fusion, transformation, electroporation, biolistics, lipofection, microinjection, or other convenient means. Where an amplifiable gene is being employed, the amplifiable gene may serve as the selection marker for selecting hosts into which the amplifiable gene has been introduced. Alternatively, one may include with the amplifiable gene another marker, such as a drug resistance marker, e.g. neomycin resistance (G418 in mammalian cells), hygromycin in resistance etc., or an auxotrophy marker (HIS3, TRP1, LEU2, URA3, ADE2, LYS2, etc.) for use in yeast cells.

[0341]Depending upon the nature of the modification and associated targeting construct, various techniques may be employed for identifying targeted integration. Conveniently, the DNA may be digested with one or more restriction enzymes and the fragments probed with an appropriate DNA fragment which will identify the properly sized restriction fragment associated with integration.

[0342]One may use different promoter sequences, enhancer sequences, or other sequence which will allow for enhanced levels of expression in the expression host. Thus, one may combine an enhancer from one source, a promoter region from another source, a 5'-noncoding region upstream from the initiation methionine from the same or different source as the other sequences and the like. One may provide for an intron in the non-coding region with appropriate splice sites or for an alternative 3'-untranslated sequence or polyadenylation site. Depending upon the particular purpose of the modification, any of these sequences may be introduced, as desired.

[0343]Where selection is intended, the sequence to be integrated will have with it a marker gene, which allows for selection. The marker gene may conveniently be downstream from the target gene and may include resistance to a cytotoxic agent, e.g. antibiotics, heavy metals, or the like, resistance or susceptibility to HAT, gancyclovir, etc., complementation to an auxotrophic host, particularly by using an auxotrophic yeast as the host for the subject manipulations, or the like. The marker gene may also be on a separate DNA molecule, particularly with primary mammalian cells. Alternatively, one may screen the various transformants, due to the high efficiency of recombination in yeast, by using hybridization analysis, PCR, sequencing, or the like.

[0344]For homologous recombination, constructs can be prepared where the amplifiable gene will be flanked, normally on both sides with DNA homologous with the DNA of the target region. Depending upon the nature of the integrating DNA and the purpose of the integration, the homologous DNA will generally be within 100 kb, usually 50 kb, preferably about 25 kb, of the transcribed region of the target gene, more preferably within 2 kb of the target gene. Where modeling of the gene is intended, homology will usually be present proximal to the site of the mutation. The homologous DNA may include the 5'-upstream region outside of the transcriptional regulatory region or comprising any enhancer sequences, transcriptional initiation sequences, adjacent sequences, or the like. The homologous region may include a portion of the coding region, where the coding region may be comprised only of an open reading frame or combination of exons and introns. The homologous region may comprise all or a portion of an intron, where all or a portion of one or more exons may also be present. Alternatively, the homologous region may comprise the 3'-region, so as to comprise all or a portion of the transcriptional termination region, or the region 3' of this region. The homologous regions may extend over all or a portion of the target gene or be outside the target gene comprising all or a portion of the transcriptional regulatory regions and/or the structural gene.

[0345]The integrating constructs may be prepared in accordance with conventional ways, where sequences may be synthesized, isolated from natural sources, manipulated, cloned, ligated, subjected to in vitro mutagenesis, primer repair, or the like. At various stages, the joined sequences may be cloned and analyzed by restriction analysis, sequencing, or the like. Usually during the preparation of a construct where various fragments are joined, the fragments, intermediate constructs and constructs will be carried on a cloning vector comprising a replication system functional in a prokaryotic host, e.g., E. coli and a marker for selection, e.g., biocide resistance, complementation to an auxotrophic host, etc. Other functional sequences may also be present, such as polylinkers, for ease of introduction and excision of the construct or portions thereof, or the like. A large number of cloning vectors are available such as pBR322, the pUC series, etc. These constructs may then be used for integration into the primary mammalian host.

[0346]In the case of the primary mammalian host, a replicating vector may be used. Usually, such vector will have a viral replication system, such as SV40, bovine papilloma virus, adenovirus, or the like. The linear DNA sequence vector may also have a selectable marker for identifying transfected cells. Selectable markers include the neo gene, allowing for selection with G418, the herpes tk gene for selection with HAT medium, the gpt gene with mycophenolic acid, complementation of an auxotrophic host, etc.

[0347]The vector may or may not be capable of stable maintenance in the host. Where the vector is capable of stable maintenance, the cells will be screened for homologous integration of the vector into the genome of the host, where various techniques for curing the cells may be employed. Where the vector is not capable of stable maintenance, for example, where a temperature sensitive replication system is employed, one may change the temperature from the permissive temperature to the non-permissive temperature, so that the cells may be cured of the vector. In this case, only those cells having integration of the construct comprising the amplifiable gene and, when present, the selectable marker, will be able to survive selection.

[0348]Where a selectable marker is present, one may select for the presence of the targeting construct by means of the selectable marker. Where the selectable marker is not present, one may select for the presence of the construct by the amplifiable gene. For the neo gene or the herpes tk gene, one could employ a medium for growth of the transformants of about 0.1-1 mg/ml of G418 or may use HAT medium, respectively. Where DHFR is the amplifiable gene, the selective medium may include from about 0.01-0.5 μM of methotrexate or be deficient in glycine-hypoxanthine-thymidine and have dialysed serum (GHT media).

[0349]The DNA can be introduced into the expression host by a variety of techniques that include calcium phosphate/DNA co-precipitates, microinjection of DNA into the nucleus, electroporation, yeast protoplast fusion with intact cells, transfection, polycations, e.g., polybrene, polyomithine, etc., or the like. The DNA may be single or double stranded DNA, linear or circular. The various techniques for transforming mammalian cells are well known (see Keown et al., Methods Enzymol. (1989); Keown et al., Methods Enzymol. 185:527-537 (1990); Mansour et al., Nature 336:348-352, (1988); all of which are herein incorporated by reference in their entirety).

[0350](d) Insect Constructs and Transformed Insect Cells

[0351]The present invention also relates to an insect recombinant vectors comprising exogenous genetic material. The present invention also relates to an insect cell comprising an insect recombinant vector. The present invention also relates to methods for obtaining a recombinant insect host cell, comprising introducing into an insect cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

[0352]The insect recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures and can bring about the expression of the nucleic acid sequence. The choice of a vector will typically depend on the compatibility of the vector with the insect host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the insect host. In addition, the insect vector may be an expression vector. Nucleic acid molecules can be suitably inserted into a replication vector for expression in the insect cell under a suitable promoter for insect cells. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid molecule to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for insect cell transformation generally include, but are not limited to, one or more of the following: a signal sequence, origin of replication, one or more marker genes and an inducible promoter.

[0353]The insect vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the insect cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the insect host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the insect host cell and, furthermore, may be non-encoding or encoding sequences.

[0354]Baculovirus expression vectors (BEVs) have become important tools for the expression of foreign genes, both for basic research and for the production of proteins with direct clinical applications in human and veterinary medicine (Doerfler, Curr. Top. Microbiol. Immunol. 131:51-68 (1968); Luckow and Summers, Bio/Technology 6:47-55 (1988a); Miller, Annual Review of Microbiol. 42:177-199 (1988); Summers, Curr. Comm. Molecular Biology, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988); all of which are herein incorporated by reference in their entirety). BEVs are recombinant insect viruses in which the coding sequence for a chosen foreign gene has been inserted behind a baculovirus promoter in place of the viral gene, e.g., polyhedrin (Smith and Summers, U.S. Pat. No. 4,745,051, the entirety of which is incorporated herein by reference).

[0355]The use of baculovirus vectors relies upon the host cells being derived from Lepidopteran insects such as Spodoptera frugiperda or Trichoplusia ni. The preferred Spodoptera frugiperda cell line is the cell line Sf9. The Spodoptera frugiperda Sf9 cell line was obtained from American Type Culture Collection (Manassas, Va.) and is assigned accession number ATCC CRL 1711 (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555 (1988), the entirety of which is herein incorporated by reference). Other insect cell systems, such as the silkworm B. mori may also be used.

[0356]The proteins expressed by the BEVs are, therefore, synthesized, modified and transported in host cells derived from Lepidopteran insects. Most of the genes that have been inserted and produced in the baculovirus expression vector system have been derived from vertebrate species. Other baculovirus genes in addition to the polyhedrin promoter may be employed to advantage in a baculovirus expression system. These include immediate-early (alpha), delayed-early (β), late (γ), or very late (delta), according to the phase of the viral infection during which they are expressed. The expression of these genes occurs sequentially, probably as the result of a "cascade" mechanism of transcriptional regulation. (Guarino and Summers, J. Virol. 57:563-571 (1986); Guarino and Summers, J. Virol. 61:2091-2099 (1987); Guarino and Summers, Virol. 162:444-451 (1988); all of which are herein incorporated by reference in their entirety).

[0357]Insect recombinant vectors are useful as intermediates for the infection or transformation of insect cell systems. For example, an insect recombinant vector containing a nucleic acid molecule encoding a baculovirus transcriptional promoter followed downstream by an insect signal DNA sequence is capable of directing the secretion of the desired biologically active protein from the insect cell. The vector may utilize a baculovirus transcriptional promoter region derived from any of the over 500 baculoviruses generally infecting insects, such as for example the Orders Lepidoptera, Diptera, Orthoptera, Coleoptera and Hymenoptera, including for example but not limited to the viral DNAs of Autographa californica MNPV, Bombyx mori NPV, Trichoplusia ni MNPV, Rachiplusia ou MNPV or Galleria mellonella MNPV, wherein said baculovirus transcriptional promoter is a baculovirus immediate-early gene IEl or IEN promoter; an immediate-early gene in combination with a baculovirus delayed-early gene promoter region selected from the group consisting of 39K and a HindIII-k fragment delayed-early gene; or a baculovirus late gene promoter. The immediate-early or delayed-early promoters can be enhanced with transcriptional enhancer elements. The insect signal DNA sequence may code for a signal peptide of a Lepidopteran adipokinetic hormone precursor or a signal peptide of the Manduca sexta adipokinetic hormone precursor (Summers, U.S. Pat. No. 5,155,037; the entirety of which is herein incorporated by reference). Other insect signal DNA sequences include a signal peptide of the Orthoptera Schistocerca gregaria locust adipokinetic hormone precursor and the Drosophila melanogaster cuticle genes CP1, CP2, CP3 or CP4 or for an insect signal peptide having substantially a similar chemical composition and function (Summers, U.S. Pat. No. 5,155,037).

[0358]Insect cells are distinctly different from animal cells. Insects have a unique life cycle and have distinct cellular properties such as the lack of intracellular plasminogen activators in which are present in vertebrate cells. Another difference is the high expression levels of protein products ranging from 1 to greater than 500 mg/liter and the ease at which cDNA can be cloned into cells (Frasier, In Vitro Cell. Dev. Biol. 25:225 (1989); Summers and Smith, In: A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555 (1988), both of which are incorporated by reference in their entirety).

[0359]Recombinant protein expression in insect cells is achieved by viral infection or stable transformation. For viral infection, the desired gene is cloned into baculovirus at the site of the wild-type polyhedron gene (Webb and Summers, Technique 2:173 (1990); Bishop and Posse, Adv. Gene Technol. 1:55 (1990); both of which are incorporated by reference in their entirety). The polyhedron gene is a component of a protein coat in occlusions which encapsulate virus particles. Deletion or insertion in the polyhedron gene results the failure to form occlusion bodies. Occlusion negative viruses are morphologically different from occlusion positive viruses and enable one skilled in the art to identify and purify recombinant viruses.

[0360]The vectors of present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. Selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, a nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the insect host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof. The promoter may be any nucleic acid sequence which shows transcriptional activity in the insect host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell.

[0361]For example, a nucleic acid molecule encoding a protein or fragment thereof may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5' terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the insect host cell of choice may be used in the present invention.

[0362]A polyadenylation sequence may also be operably linked to the 3' terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the insect host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention.

[0363]To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed polypeptide within the cell, it is preferred that expression of the polypeptide gene gives rise to a product secreted outside the cell. To this end, the protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the insect host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5' end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof.

[0364]At present, a mode of achieving secretion of a foreign gene product in insect cells is by way of the foreign gene's native signal peptide. Because the foreign genes are usually from non-insect organisms, their signal sequences may be poorly recognized by insect cells and hence, levels of expression may be suboptimal. However, the efficiency of expression of foreign gene products seems to depend primarily on the characteristics of the foreign protein. On average, nuclear localized or non-structural proteins are most highly expressed, secreted proteins are intermediate and integral membrane proteins are the least expressed. One factor generally affecting the efficiency of the production of foreign gene products in a heterologous host system is the presence of native signal sequences (also termed presequences, targeting signals, or leader sequences) associated with the foreign gene. The signal sequence is generally coded by a DNA sequence immediately following (5' to 3') the translation start site of the desired foreign gene.

[0365]The expression dependence on the type of signal sequence associated with a gene product can be represented by the following example: If a foreign gene is inserted at a site downstream from the translational start site of the baculovirus polyhedrin gene so as to produce a fusion protein (containing the N-terminus of the polyhedrin structural gene), the fused gene is highly expressed. But less expression is achieved when a foreign gene is inserted in a baculovirus expression vector immediately following the transcriptional start site and totally replacing the polyhedrin structural gene.

[0366]Insertions into the region -50 to -1 significantly alter (reduce) steady state transcription which, in turn, reduces translation of the foreign gene product. Use of the pVL941 vector optimizes transcription of foreign genes to the level of the polyhedrin gene transcription. Even though the transcription of a foreign gene may be optimal, optimal translation may vary because of several factors involving processing: signal peptide recognition, mRNA and ribosome binding, glycosylation, disulfide bond formation, sugar processing, oligomerization, for example.

[0367]The properties of the insect signal peptide are expected to be more optimal for the efficiency of the translation process in insect cells than those from vertebrate proteins. This phenomenon can generally be explained by the fact that proteins secreted from cells are synthesized as precursor molecules containing hydrophobic N-terminal signal peptides. The signal peptides direct transport of the select protein to its target membrane and are then cleaved by a peptidase on the membrane, such as the endoplasmic reticulum, when the protein passes through it.

[0368]Another exemplary insect signal sequence is the sequence encoding for Drosophila cuticle proteins such as CP1, CP2, CP3 or CP4 (Summers, U.S. Pat. No. 5,278,050; the entirety of which is herein incorporated by reference). Most of a 9 kb region of the Drosophila genome containing genes for the cuticle proteins has been sequenced. Four of the five cuticle genes contains a signal peptide coding sequence interrupted by a short intervening sequence (about 60 base pairs) at a conserved site. Conserved sequences occur in the 5' mRNA untranslated region, in the adjacent 35 base pairs of upstream flanking sequence and at -200 base pairs from the mRNA start position in each of the cuticle genes.

[0369]Standard methods of insect cell culture, cotransfection and preparation of plasmids are set forth in Summers and Smith (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Agricultural Experiment Station Bulletin No. 1555, Texas A&M University (1987)). Procedures for the cultivation of viruses and cells are described in Volkman and Summers, J. Virol. 19:820-832 (1975) and Volkman et al., J. Virol. 19:820-832 (1976); both of which are herein incorporated by reference in their entirety.

[0370](e) Bacterial Constructs and Transformed Bacterial Cells

[0371]The present invention also relates to a bacterial recombinant vector comprising exogenous genetic material. The present invention also relates to a bacteria cell comprising a bacterial recombinant vector. The present invention also relates to methods for obtaining a recombinant bacteria host cell, comprising introducing into a bacterial host cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

[0372]The bacterial recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the bacterial host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the bacterial host. In addition, the bacterial vector may be an expression vector. Nucleic acid molecules encoding protein homologues or fragments thereof can, for example, be suitably inserted into a replicable vector for expression in the bacterium under the control of a suitable promoter for bacteria. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for bacterial transformation generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes and an inducible promoter.

[0373]In general, plasmid vectors containing replicon and control sequences that are derived from species compatible with the host cell are used in connection with bacterial hosts. The vector ordinarily carries a replication site, as well as marking sequences that are capable of providing phenotypic selection in transformed cells. For example, E. coli is typically transformed using pBR322, a plasmid derived from an E. coli species (see, e.g., Bolivar et al., Gene 2:95 (1977); the entirety of which is herein incorporated by reference). pBR322 contains genes for ampicillin and tetracycline resistance and thus provides easy means for identifying transformed cells. The pBR322 plasmid, or other microbial plasmid or phage, also generally contains, or is modified to contain, promoters that can be used by the microbial organism for expression of the selectable marker genes.

[0374]Nucleic acid molecules encoding protein or fragments thereof may be expressed not only directly, but also as a fusion with another polypeptide, preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature polypeptide. In general, the signal sequence may be a component of the vector, or it may be a part of the polypeptide DNA that is inserted into the vector. The heterologous signal sequence selected should be one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. For bacterial host cells that do not recognize and process the native polypeptide signal sequence, the signal sequence is substituted by a bacterial signal sequence selected, for example, from the group consisting of the alkaline phosphatase, penicillinase, lpp, or heat-stable enterotoxin II leaders.

[0375]Both expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors this sequence is one that enables the vector to replicate independently of the host chromosomal DNA and includes origins of replication or autonomously replicating sequences. Such sequences are well known for a variety of bacteria. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria.

[0376]Expression and cloning vectors also generally contain a selection gene, also termed a selectable marker. This gene encodes a protein necessary for the survival or growth of transformed host cells grown in a selective culture medium. Host cells not transformed with the vector containing the selection gene will not survive in the culture medium. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous protein homologue or fragment thereof produce a protein conferring drug resistance and thus survive the selection regimen.

[0377]The expression vector for producing a protein or fragment thereof can also contains an inducible promoter that is recognized by the host bacterial organism and is operably linked to the nucleic acid encoding, for example, the nucleic acid molecule encoding the protein homologue or fragment thereof of interest. Inducible promoters suitable for use with bacterial hosts include the β-lactamase and lactose promoter systems (Chang et al., Nature 275:615 (1978); Goeddel et al., Nature 281:544 (1979); both of which are herein incorporated by reference in their entirety), the arabinose promoter system (Guzman et al., J. Bacteriol. 174:7716-7728 (1992); the entirety of which is herein incorporated by reference), alkaline phosphatase, a tryptophan (trp) promoter system (Goeddel, Nucleic Acids Res. 8:4057 (1980); EP 36,776; both of which are herein incorporated by reference in their entirety) and hybrid promoters such as the tac promoter (deBoer et al., Proc. Natl. Acad. Sci. (USA) 80:21-25 (1983); the entirety of which is herein incorporated by reference). However, other known bacterial inducible promoters are suitable (Siebenlist et al., Cell 20:269 (1980); the entirety of which is herein incorporated by reference).

[0378]Promoters for use in bacterial systems also generally contain a Shine-Dalgamo (S.D.) sequence operably linked to the DNA encoding the polypeptide of interest. The promoter can be removed from the bacterial source DNA by restriction enzyme digestion and inserted into the vector containing the desired DNA.

[0379]Construction of suitable vectors containing one or more of the above-listed components employs standard ligation techniques. Isolated plasmids or DNA fragments are cleaved, tailored and re-ligated in the form desired to generate the plasmids required. Examples of available bacterial expression vectors include, but are not limited to, the multifunctional E. coli cloning and expression vectors such as Bluescript® (Stratagene, La Jolla, Calif.), in which, for example, encoding an A. nidulans protein homologue or fragment thereof homologue, may be ligated into the vector in frame with sequences for the amino-terminal Met and the subsequent 7 residues of β-galactosidase so that a hybrid protein is produced; pIN vectors (Van Heeke and Schuster, J. Biol. Chem. 264:5503-5509 (1989), the entirety of which is herein incorporated by reference); and the like. pGEX vectors (Promega, Madison Wis. U.S.A.) may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems are designed to include heparin, thrombin or factor XA protease cleavage sites so that the cloned polypeptide of interest can be released from the GST moiety at will.

[0380]Suitable host bacteria for a bacterial vector include archaebacteria and eubacteria, especially eubacteria and most preferably Enterobacteriaceae. Examples of useful bacteria include Escherichia, Enterobacter, Azotobacter, Erwin ia, Bacillus, Pseudomonas, Klebsiella, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla and Paracoccus. Suitable E. coli hosts include E. coli W3110 (American Type Culture Collection (ATCC) 27,325, Manassas, Va. U.S.A.), E. coli 294 (ATCC 31,446), E. coli B and E. coli X1776 (ATCC 31,537). These examples are illustrative rather than limiting. Mutant cells of any of the above-mentioned bacteria may also be employed. It is, of course, necessary to select the appropriate bacteria taking into consideration replicability of the replicon in the cells of a bacterium. For example, E. coli, Serratia, or Salmonella species can be suitably used as the host when well known plasmids such as pBR322, pBR325, pACYC177, or pKN410 are used to supply the replicon. E. coli strain W3110 is a preferred host or parent host because it is a common host strain for recombinant DNA product fermentations. Preferably, the host cell should secrete minimal amounts of proteolytic enzymes.

[0381]Host cells are transfected and preferably transformed with the above-described vectors and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.

[0382]Numerous methods of transfection are known to the ordinarily skilled artisan, for example, calcium phosphate and electroporation. Depending on the host cell used, transformation is done using standard techniques appropriate to such cells. The calcium treatment employing calcium chloride, as described in section 1.82 of Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989), is generally used for bacterial cells that contain substantial cell-wall barriers. Another method for transformation employs polyethylene glycol/DMSO, as described in Chung and Miller (Chung and Miller, Nucleic Acids Res. 16:3580 (1988); the entirety of which is herein incorporated by reference). Yet another method is the use of the technique termed electroporation.

[0383]Bacterial cells used to produce the polypeptide of interest for purposes of this invention are cultured in suitable media in which the promoters for the nucleic acid encoding the heterologous polypeptide can be artificially induced as described generally, e.g., in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989). Examples of suitable media are given in U.S. Pat. Nos. 5,304,472 and 5,342,763; both of which are incorporated by reference in their entirety.

[0384]In addition to the above discussed procedures, practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of macromolecules (e.g., DNA molecules, plasmids, etc.), generation of recombinant organisms and the screening and isolating of clones, (see for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989); Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995), the entirety of which is herein incorporated by reference; Birren et al., Genome Analysis: Analyzing DNA, 1, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference).

[0385](f) Computer Readable Media

[0386]The nucleotide sequence provided in SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof, or complement thereof, or a nucleotide sequence at least 90% identical, preferably 95%, identical even more preferably 99% or 100% identical to the sequence provided in SEQ ID NO: 1 through SEQ ID NO: 677 or fragment thereof, or complement thereof, can be "provided" in a variety of mediums to facilitate use. Such a medium can also provide a subset thereof in a form that allows a skilled artisan to examine the sequences.

[0387]A preferred subset of nucleotide sequences are those nucleic acid sequences that encode a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof or fragment of either.

[0388]A further preferred subset of nucleic acid sequences is where the subset of sequences is two proteins or fragments thereof, more preferably three proteins or fragments thereof and even more preferable four transcription factors or fragments thereof, these nucleic acid sequences are selected from the group that comprises a nucleic acid molecule that encodes a putative maize or soybean chlorophyll synthetase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize or soybean protochlorophyllide reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean coproporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean protoporphyrinogen oxidase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a putative maize uroporphyrinogen decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean porphobilinogen synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean hydroxymethylbilane synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate-1-semialdehyde 2,1-aminomutase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamate tRNA ligase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean glutamyl-tRNA reductase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean Mg-chelatase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or soybean ferrochelatase enzyme or complement thereof or fragment of either.

[0389]In one application of this embodiment, a nucleotide sequence of the present invention can be recorded on computer readable media. As used herein, "computer readable media" refers to any medium that can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc, storage medium and magnetic tape: optical storage media such as CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media. A skilled artisan can readily appreciate how any of the presently known computer readable mediums can be used to create a manufacture comprising computer readable medium having recorded thereon a nucleotide sequence of the present invention.

[0390]As used herein, "recorded" refers to a process for storing information on computer readable medium. A skilled artisan can readily adopt any of the presently known methods for recording information on computer readable medium to generate media comprising the nucleotide sequence information of the present invention. A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon a nucleotide sequence of the present invention. The choice of the data storage structure will generally be based on the means chosen to access the stored information. In addition, a variety of data processor programs and formats can be used to store the nucleotide sequence information of the present invention on computer readable medium. The sequence information can be represented in a word processing text file, formatted in commercially-available software such as WordPerfect and Microsoft Word, or represented in the form of an ASCII file, stored in a database application, such as DB2, Sybase, Oracle, or the like. A skilled artisan can readily adapt any number of data processor structuring formats (e.g. text file or database) in order to obtain computer readable medium having recorded thereon the nucleotide sequence information of the present invention.

[0391]By providing one or more of nucleotide sequences of the present invention, a skilled artisan can routinely access the sequence information for a variety of purposes. Computer software is publicly available which allows a skilled artisan to access sequence information provided in a computer readable medium. The examples which follow demonstrate how software which implements the BLAST (Altschul et al., J. Mol. Biol. 215:403-410 (1990), the entirety of which is herein incorporated by reference) and BLAZE (Brutlag et al., Comp. Chem. 17:203-207 (1993), the entirety of which is herein incorporated by reference) search algorithms on a Sybase system can be used to identify open reading frames (ORFs) within the genome that contain homology to ORFs or proteins from other organisms. Such ORFs are protein-encoding fragments within the sequences of the present invention and are useful in producing commercially important proteins such as enzymes used in amino acid biosynthesis, metabolism, transcription, translation, RNA processing, nucleic acid and a protein degradation, protein modification and DNA replication, restriction, modification, recombination and repair.

[0392]The present invention further provides systems, particularly computer-based systems, which contain the sequence information described herein. Such systems are designed to identify commercially important fragments of the nucleic acid molecule of the present invention. As used herein, "a computer-based system" refers to the hardware means, software means and data storage means used to analyze the nucleotide sequence information of the present invention. The minimum hardware means of the computer-based systems of the present invention comprises a central processing unit (CPU), input means, output means and data storage means. A skilled artisan can readily appreciate that any one of the currently available computer-based system are suitable for use in the present invention.

[0393]As indicated above, the computer-based systems of the present invention comprise a data storage means having stored therein a nucleotide sequence of the present invention and the necessary hardware means and software means for supporting and implementing a search means. As used herein, "data storage means" refers to memory that can store nucleotide sequence information of the present invention, or a memory access means which can access manufactures having recorded thereon the nucleotide sequence information of the present invention. As used herein, "search means" refers to one or more programs which are implemented on the computer-based system to compare a target sequence or target structural motif with the sequence information stored within the data storage means. Search means are used to identify fragments or regions of the sequence of the present invention that match a particular target sequence or target motif. A variety of known algorithms are disclosed publicly and a variety of commercially available software for conducting search means are available can be used in the computer-based systems of the present invention. Examples of such software include, but are not limited to, MacPattern (EMBL), BLASTIN and BLASTIX (NCBIA). One of the available algorithms or implementing software packages for conducting homology searches can be adapted for use in the present computer-based systems.

[0394]The most preferred sequence length of a target sequence is from about 10 to 100 amino acids or from about 30 to 300 nucleotide residues. However, it is well recognized that during searches for commercially important fragments of the nucleic acid molecules of the present invention, such as sequence fragments involved in gene expression and protein processing, may be of shorter length.

[0395]As used herein, "a target structural motif," or "target motif," refers to any rationally selected sequence or combination of sequences in which the sequences the sequence(s) are chosen based on a three-dimensional configuration which is formed upon the folding of the target motif. There are a variety of target motifs known in the art. Protein target motifs include, but are not limited to, enzymatic active sites and signal sequences. Nucleic acid target motifs include, but are not limited to, promoter sequences, cis elements, hairpin structures and inducible expression elements (protein binding sequences).

[0396]Thus, the present invention further provides an input means for receiving a target sequence, a data storage means for storing the target sequences of the present invention sequence identified using a search means as described above and an output means for outputting the identified homologous sequences. A variety of structural formats for the input and output means can be used to input and output information in the computer-based systems of the present invention. A preferred format for an output means ranks fragments of the sequence of the present invention by varying degrees of homology to the target sequence or target motif Such presentation provides a skilled artisan with a ranking of sequences which contain various amounts of the target sequence or target motif and identifies the degree of homology contained in the identified fragment.

[0397]A variety of comparing means can be used to compare a target sequence or target motif with the data storage means to identify sequence fragments sequence of the present invention. For example, implementing software which implement the BLAST and BLAZE algorithms (Altschul et al., J. Mol. Biol. 215:403-410 (1990)) can be used to identify open frames within the nucleic acid molecules of the present invention. A skilled artisan can readily recognize that any one of the publicly available homology search programs can be used as the search means for the computer-based systems of the present invention.

[0398]Having now generally described the invention, the same will be more readily understood through reference to the following examples which are provided by way of illustration and are not intended to be limiting of the present invention, unless specified.

Example 1

[0399]The MONN01 cDNA library is a normalized library generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0400]The SATMON001 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) immature tassels at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0401]The SATMON003 library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) roots at the V6 developmental stage. Seeds are planted at a depth of approximately 3 cm in coil into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, the seedlings are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting at a concentration of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in approximately 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6 leaf development stage. The root system is cut from maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0402]The SATMON004 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0403]The SATMON005 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign Ill., U.S.A.) root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at -80° C. until RNA preparation.

[0404]The SATMON006 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign Ill., U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0405]The SATMON007 cDNA library is generated from the primary root tissue of 5 day old maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). After germination, the trays, along with the moist paper, are moved to a greenhouse where the maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles for approximately 5 days. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. The primary root tissue is collected when the seedlings are 5 days old. At this stage, the primary root (radicle) is pushed through the coleorhiza which itself is pushed through the seed coat. The primary root, which is about 2-3 cm long, is cut and immediately frozen in liquid nitrogen and then stored at -80° C. until RNA preparation.

[0406]The SATMON008 cDNA library is generated from the primary shoot (coleoptile 2-3 cm) of maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings which are approximately 5 days old. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to a greenhouse at 15 hr daytime/9 hr nighttime cycles and grown until they are 5 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 5 days old. At this stage, the primary shoot (coleoptile) is pushed through the seed coat and is about 2-3 cm long. The coleoptile is dissected away from the rest of the seedling, immediately frozen in liquid nitrogen and then stored at -80° C. until RNA preparation.

[0407]The SATMON009 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves at the 8 leaf stage (V8 plant development stage). Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 8-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical, are cut at the base of the leaves. The leaves are then pooled and then immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0408]The SATMON010 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the V8 development stage. The root system is cut from this mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0409]The SATMON011 cDNA library is generated from undeveloped maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The second youngest leaf which is at the base of the apical leaf of V6 stage maize plant is cut at the base and immediately transferred to liquid nitrogen containers in which the leaf is crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0410]The SATMON012 cDNA library is generated from 2 day post germination maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to the greenhouse and grown at 15 hr daytime/9 hr nighttime cycles until 2 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 2 days old. At the two day stage, the coleorhiza is pushed through the seed coat and the primary root (the radicle) is pierced the coleorhiza but is barely visible. Also, at this two day stage, the coleoptile is just emerging from the seed coat. The 2 days post germination seedlings are then immersed in liquid nitrogen and crushed. The harvested tissue is stored at -80° C. until preparation of total RNA.

[0411]The SATMON013 cDNA library is generated from apical maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) meristem founder at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, the plant is at the 4 leaf stage. The lead at the apex of the V4 stage maize plant is referred to as the meristem founder. This apical meristem founder is cut, immediately frozen in liquid nitrogen and crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0412]The SATMON014 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm fourteen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the maize plant ear shoots are ready for fertilization. At this stage, the ear shoots are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are pollinated and 14 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at -80° C. until RNA preparation.

[0413]The SATMON016 library is a maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) sheath library collected at the V8 developmental stage. Seeds are planted in a depth of approximately 3 cm in solid into 2-3 inch pots containing Metro growing medium. After 2-3 weeks growth, they are transplanted into 10'' pots containing the same. Plants are watered daily before transplantation and approximately the times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. When the maize plants are at the V8 stage the 5^th and 6^th leaves from the bottom exhibit fully developed leaf blades. At the base of these leaves, the ligule is differentiated and the leaf blade is joined to the sheath. The sheath is dissected away from the base of the leaf then the sheath is frozen in liquid nitrogen and crushed. The tissue is then stored at -80° C. until RNA preparation.

[0414]The SATMON017 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo seventeen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth the seeds are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are fertilized and 21 days after pollination, the ears are pulled out and the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at -80° C. until RNA preparation.

[0415]The SATMON019 (Lib3054) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) culm (stem) at the V8 developmental stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. When the maize plant is at the V8 stage, the 5th and 6th leaves from the bottom have fully developed leaf blades. The region between the nodes of the 5th and the sixth leaves from the bottom is the region of the stem that is collected. The leaves are pulled out and the sheath is also torn away from the stem. This stem tissue is completely free of any leaf and sheath tissue. The stem tissue is then frozen in liquid nitrogen and stored at -80° C. until RNA preparation.

[0416]The SATMON020 cDNA library is from a maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Initiated Callus. Petri plates containing approximately 25 ml of Type II initiation media are prepared. This medium contains N6 salts and vitamins, 3% sucrose, 2.3 g/liter proline 0. 1 g/liter enzymatic casein hydrolysate, 2 mg/liter 2,4-dichloro phenoxy-acetic acid (2,4, D), 15.3 mg/liter AgNO₃ and 0.8% bacto agar and is adjusted to pH 6.0 before autoclaving. At 9-11 days after pollination, an ear with immature embryos measuring approximately 1-2 mm in length is chosen. The husks and silks are removed and then the ear is broken into halves and placed in an autoclaved solution of Clorox/TWEEN 20 sterilizing solution. Then the ear is rinsed with deionized water. Then each embryo is extracted from the kernel. Intact embryos are placed in contact with the medium, scutellar side up). Multiple embryos are plated on each plate and the plates are incubated in the dark at 25° C. Type II calluses are friable, can be subcultured with a spatula, frequently regenerate via somatic embryogenesis and are relatively undifferentiated. As seen in the microscope, the Tape II calluses show color ranging from translucent to light yellow and heterogeneity on with respect to embryoid structure as well as stage of embryoid development. Once Type II callus are formed, the calluses is transferred to type II callus maintenance medium without AgN0₃. Every 7-10 days, the callus is subcultured. About 4 weeks after embryo isolation the callus is removed from the plates and then frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0417]The SATMON021 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb Ill., U.S.A.) tassel at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. As the maize plant enters the V8 stage, tassels which are 15-20 cm in length are collected and frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0418]The SATMON022 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) ear (growing silks) at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the plant is in the V8 stage. At this stage, some immature ear shoots are visible. The immature ear shoots (approximately 1 cm in length) are pulled out, frozen in liquid nitrogen and then stored at -80° C. until RNA preparation.

[0419]The SATMON23 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) car (growing silk) at the V8 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. When the tissue is harvested at the V8 stage, the length of the ear that is harvested is about 10-15 cm and the silks are just exposed (approximately 1 inch). The ear along with the silks is frozen in liquid nitrogen and then the tissue is stored at -80° C. until RNA preparation.

[0420]The SATMON024 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) tassel at the V9 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. As a maize plant enters the V9 stage, the tassel is rapidly developing and a 37 cm tassel along with the glume, anthers and pollen is collected and frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0421]The SATMON025 cDNA library is from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Regenerated Callus. Type II callus is grown in initiation media as described for SATMON020 and then the embryoids on the surface of the Type II callus are allowed to mature and germinate. The 1-2 gm fresh weight of the soft friable type callus containing numerous embryoids are transferred to 100×15 mm petri plates containing 25 ml of regeneration media. Regeneration media consists of Murashige and Skoog (MS) basal salts, modified White's vitamins (0.2 g/liter glycine and 0.5 g/liter myo-inositoland 0.8% bacto agar (6SMS0D)). The plates are then placed in the dark after covering with parafilm. After 1 week, the plates are moved to a lighted growth chamber with 16 hr light and 8 hr dark photoperiod. Three weeks after plating the Type II callus to 6SMS0D, the callus exhibit shoot formation. The callus and the shoots are transferred to fresh 6SMS0D plates for another 2 weeks. The callus and the shoots are then transferred to petri plates with reduced sucrose (3SMS0D). Upon distinct formation of a root and shoot, the newly developed green plants are then removed out with a spatula and frozen in liquid nitrogen containers. The harvested tissue is then stored at -80° C. until RNA preparation.

[0422]The SATMON026 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) juvenile/adult shift leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plants are at the 8-leaf development stage. Leaves are founded sequentially around the meristem over weeks of time and the older, more juvenile leaves arise earlier and in a more basal position than the younger, more adult leaves, which are in a more apical position. In a V8 plant, some leaves which are in the middle portion of the plant exhibit characteristics of both juvenile as well as adult leaves. They exhibit a yellowing color but also exhibit, in part, a green color. These leaves are termed juvenile/adult shift leaves. The juvenile/adult shift leaves (the 4th, 5th leaves from the bottom) are cut at the base, pooled and transferred to liquid nitrogen in which they are then crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0423]The SATMON027 cDNA library is generated from 6 day maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical, are all cut at the base of the leaves. All the leaves exhibit significant wilting. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0424]The SATMON028 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) roots at the V8 developmental stage that are subject to six days water stress. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The root system is cut, shaken and washed to remove soil. Root tissue is then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0425]The SATMON029 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings at the etiolated stage. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark for 4 days at approximately 70° F. Tissue is collected when the seedlings are 4 days old. By 4 days, the primary root has penetrated the coleorhiza and is about 4-5 cm and the secondary lateral roots have also made their appearance. The coleoptile has also pushed through the seed coat and is about 4-5 cm long. The seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0426]The SATMON030 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, they are transplanted into 10 inch pots containing the same. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant, from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 sodium vapor lamps. Tissue is collected when the maize plant is at the 4 leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0427]The SATMON031 cDNA library is generated from the maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 4-leaf development stage. The third leaf from the bottom is cut at the base and immediately frozen in liquid nitrogen and crushed. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0428]The SATMON033 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo tissue 13 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 13 days after pollination, the ears are pulled out and then the kernels are plucked cut of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at -80° C. until RNA preparation.

[0429]The SATMON034 cDNA library is generated from cold stressed maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept on at 10° C. for 7 days. After 7 days, the temperature is shifted to 15° C. for one day until germination of the seed. Tissue is collected once the seedlings are 1 day old. At this point, the coleorhiza has just pushed out of the seed coat and the primary root is just making its appearance. The coleoptile has not yet pushed completely through the seed coat and is also just making its appearance. These 1 day old cold stressed seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0430]The SATMON˜001 (Lib36, Lib83, Lib84) cDNA library is generated from maize leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V8 stage. The older more juvenile leaves in a basal position was well as the younger more adult leaves which are more apical are all cut at the base, pooled and frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0431]The SATMONN01 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Ill. U.S.A.)normalized immature tassels at the V6 plant development stage normalized tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

[0432]The SATMONN04 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at -80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

[0433]The SATMONN05 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign Ill., U.S.A.) normalized root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at -80° C. until RNA preparation. The single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

[0434]The SATMONN06 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign Ill., U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at -80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

[0435]The CMZO29 (SATMON036) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm 22 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 22 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the alurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at -80° C. until RNA preparation.

[0436]The CMz030 (Lib 143) cDNA library is generated from maize seedling tissue two days post germination. Seeds are planted on a moist filter paper on a covered try that is keep in the dark until germination. The trays are then moved to the bench top at 15 hr daytime/9 hr nighttime cycles for 2 days post-germination. The day time temperature is 80° F. and the nighttime temperature is 70° F. Tissue is collected when the seedlings are 2 days old. At this stage, the colehrhiza has pushed through the seed coat and the primary root (the radicle) is just piercing the colehrhiza and is barely visible. The seedlings are placed at 42° C. for 1 hour. Following the heat shock treatment, the seedlings are immersed in liquid nitrogen and crushed. The harvested tissue is stored at -80° C. until RNA preparation.

[0437]The CMz031 (Lib148) cDNA library is generated from maize pollen tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag to withhold pollen. Twenty-one days after pollination, prior to removing the ears, the paper bag is shaken to collect the mature pollen. The mature pollen is immediately frozen in liquid nitrogen containers and the pollen is crushed. The harvested tissue is then stored at -80° C. until RNA preparation.

[0438]The CMz033 (Lib189) cDNA library is generated from maize pooled leaf tissue. Samples are harvested from open pollinated plants. Tissue is collected from maize leaves at the anthesis stage. The leaves are collect from 10-12 plants and frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0439]The CMz034 (Lib3060) cDNA library is generated from maize mature tissue at 40 days post pollination plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from leaves located two leaves below the ear leaf. This sample represents those genes expressed during onset and early stages of leaf senescence. The leaves are pooled and immediately transferred to liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0440]The CMz035 (Lib3061) cDNA library is generated from maize endosperm tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag prior to silk emergence to withhold pollen. Thirty-two days after pollination, the ears are pulled out and the kernels are removed from the cob. Each kernel is dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the endosperms are immediately transferred to liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0441]The CMz036 (Lib3062) cDNA library is generated from maize husk tissue at the 8 week old plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from 8 week old plants. The husk is separated from the ear and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at -80° C. until RNA preparation.

[0442]The CMz037 (Lib3059) cDNA library is generated from maize pooled kernel at 12-15 days after pollination plant development stage. Sample were collected from field grown material. Whole kernels from hand pollinated (control pollination) are harvested as whole ears and immediately frozen on dry ice. Kernels from 10-12 ears were pooled and ground together in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0443]The CMz039 (Lib3066) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0444]The CMz040 (Lib3067) cDNA library is generated from maize kernel tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold pollen. Five to eight days after controlled pollination. The ears are pulled and the kernels removed. The kernels are immediately frozen in liquid nitrogen. The harvested kernels tissue is then stored at -80° C. until RNA preparation. This sample represents gene expressed in early kernel development, during periods of cell division, amyloplast biogenesis and early carbon flow across the material to filial tissue.

[0445]The CMz041 (Lib3068) cDNA library is generated from maize pollen germinating silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants when the ear shoots are ready for fertilization at the silk emergence stage. The emerging silks are pollinated with an excess of pollen under controlled pollination conditions in the green house. Eighteen hours after pollination the silks are removed from the ears and immediately frozen in liquid nitrogen containers. This sample represents genes expressed in both pollen and silk tissue early in pollination. The harvested tissue is then stored at -80° C. until RNA preparation.

[0446]The CMz042 (Lib3069) cDNA library is generated from maize ear tissue excessively pollinated at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants and the ear shoots which are ready for fertilization are at the silk emergence stage. The immature ears are pollinated with an excess of pollen under controlled pollination conditions. Eighteen hours post-pollination, the ears are removed and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at -80° C. until RNA preparation.

[0447]The CMz044 (Lib3075) cDNA library is generated from maize microspore tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature anthers from 7 week old tassels. The immature anthers are first dissected from the 7 week old tassel with a scalpel on a glass slide covered with water. The microspores (immature pollen) are released into the water and are recovered by centrifugation. The microspore suspension is immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0448]The CMz045 (Lib3076) cDNA library is generated from maize immature ear megaspore tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature ear (megaspore) obtained from 7 week old plants. The immature ears are harvested from the 7 week old plants and are approximately 2.5 to 3 cm in length. The kernels are removed from the cob immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0449]The CMz047 (Lib3078) cDNA library is generated from maize CO₂ treated high-exposure shoot tissue at the V10+ plant development stage. RX601 maize seeds are sterilized for i minute with a 10% clorox solution. The seeds are rolled in germination paper, and germinated in 0.5 mM calcium sulfate solution for two days ate 30° C. The seedlings are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium at a rate of 2-3 seedlings per pot. Twenty pots are placed into a high CO₂ environment (approximately 1000 ppm CO₂). Twenty plants were grown under ambient greenhouse CO₂ (approximately 450 ppm CO₂). Plants are watered daily before transplantation and three times a week after transplantation. Peters 20-20-20 fertilizer is also lightly applied. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. At ten days post planting, the shoots from both atmosphere are frozen in liquid nitrogen and lightly ground. The roots are washed in deionized water to remove the support media and the tissue is immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at -80° C. until RNA preparation.

[0450]The CMz048 (Lib3079) cDNA library is generated from maize basal endosperm transfer layer tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ maize plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag prior to silk emergence, to withhold the pollen. Kernels are harvested at 12 days post-pollination and placed on wet ice for dissection. The kernels are cross sectioned laterally, dissecting just above the pedicel region, including 1-2 mm of the lower endosperm and the basal endosperm transfer region. The pedicel and lower endosperm region containing the basal endosperm transfer layer is pooled and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0451]The CMz049(Lib3088) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately transferred to liquid nitrogen container. The harvested tissue is then stored at -80° C. until RNA preparation.

[0452]The CMz050 (Lib3114) cDNA library is generated from maize silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is beyond the 10-leaf development stage and the ear shoots are approximately 15-20 cm in length. The ears are pulled and silks are separated from the ears and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at -80° C. until RNA preparation.

[0453]The SOYMON001 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) total leaf tissue at the V4 plant development stage. Leaf tissue from 38, field grown V4 stage plants is harvested from the 4^th node. Leaf tissue is removed from the plants and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0454]The SOYMON002 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue at the V4 plant development stage. Root tissue from 76, field grown V4 stage plants is harvested. The root systems is cut from the soybean plant and washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0455]The SOYMON003 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0456]The SOYMON004 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledon tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0457]The SOYMON005 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after the start of imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post imbibition. At the 6 hours after imbibition stage, not all cotyledons have become fully hydrated and germination, or radicle protrusion, has not occurred. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0458]The SOYMON006 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledons tissue harvest 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post-imbibition. At the 6 hours after imbibition, not all cotyledons have become fully hydrated and germination or radicle protrusion, have not occurred. The seedlings are washed in water to remove soil, cotyledon harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0459]The SOYMON007 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days post-flowering. Seed pods from field grown plants are harvested 25 and 35 days after flowering and the seeds extracted from the pods. Approximately 4.4 g and 19.3 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0460]The SOYMON008 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested from 25 and 35 days post-flowering plants. Total leaf tissue is harvested from field grown plants. Approximately 19 g and 29 g of leaves are harvested from the fourth node of the plant 25 and 35 days post-flowering and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0461]The SOYMON009 cDNA library is generated from soybean cultivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) pod and seed tissue harvested 15 days post-flowering. Pods from field grown plants are harvested 15 days post-flowering. Approximately 3 g of pod tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0462]The SOYMON010 cDNA library is generated from soybean cultivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) seed tissue harvested 40 days post-flowering. Pods from field grown plants are harvested 40 days post-flowering. Pods and seeds are separated, approximately 19 g of seed tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0463]The SOYMON011 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4 h node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0464]The SOYMON012 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue. Leaves from field grown plants are harvested from the fourth node 15 days post-flowering. Approximately 12 g of leaves are harvested and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0465]The SOYMON013 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root and nodule tissue. Approximately, 28 g of root tissue from field grown plants is harvested 15 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0466]The SOYMON014 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days after flowering. Seed pods from field grown plants are harvested 15 days after flowering and the seeds extracted from the pods. Approximately 5 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0467]The SOYMON015 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 45 and 55 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 19 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0468]The SOYMON016 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately, 61 g and 38 g of root tissue from field grown plants is harvested 25 and 35 days post-flowering is harvested. The root system is cut from the soybean plant and washed with water to free it from the soil. The tissue is placed in 14 ml polystyrene tubes and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0469]The SOYMON017 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately 28 g of root tissue from field grown plants is harvested 45 and 55 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0470]The SOYMON018 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 45 and 55 days post-flowering. Leaves from field grown plants are harvested 45 and 55 days after flowering from the fourth node. Approximately 27 g and 33 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0471]The SOYMON019 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0472]The SOYMON020 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 65 and 75 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 14 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0473]The SOYMON021 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Plants are grown in tissue culture at room temperature. At approximately 6 weeks post-germination, the plants are exposed to sterilized Soybean Cyst Nematode eggs. Infection is then allowed to progress for 10 days. After the 10 day infection process, the tissue is harvested. Agar from the culture medium and nematodes are removed and the root tissue is immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0474]The SOYMON022 (Lib3030) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) partially opened flower tissue. Partially to fully opened flower tissue is harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. A total of 3 g of flower tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0475]The SOYMON023 cDNA library is generated from soybean genotype BW211S Null (Tohoku University, Morioka, Japan) seed tissue harvested 15 and 40 days post-flowering. Seed pods from field grown plants are harvested 15 and 40 days post-flowering and the seeds extracted from the pods. Approximately 0.7 g and 142 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0476]The SOYMON024 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) internode-2 tissue harvested 18 days post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. The plants are grown in a greenhouse for 18 days after the start of imbibition at ambient temperature. Soil is checked and watered daily to maintain even moisture conditions. Stem tissue is harvested 18 days after the start of imbibition. The samples are divided into hypocotyl and internodes 1 through 5. The fifth internode contains some leaf bud material. Approximately 3 g of each sample is harvested and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0477]The SOYMON025 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 65 days post-flowering. Leaves are harvested from the fourth node of field grown plants 65 days post-flowering. Approximately 18.4 g of leaf tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0478]SOYMON026 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue harvested 65 and 75 days post-flowering. Approximately 27 g and 40 g of root tissue from field grown plants is harvested 65 and 75 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0479]The SOYMON027 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 days post-flowering. Seed pods from field grown plants are harvested 25 days post-flowering and the seeds extracted from the pods. Approximately 17 g of seeds are harvested from the seed pods and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0480]The SOYMON028 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed root tissue. The plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of development, water is withheld from half of the plant collection (drought stressed population). After 3 days, half of the plants from the drought stressed condition and half of the plants from the control population are harvested. After another 3 days (6 days post drought induction) the remaining plants are harvested. A total of 27 g and 40 g of root tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0481]The SOYMON029 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar PI07354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Late fall to early winter greenhouse grown plants are exposed to Soybean Cyst Nematode eggs. At 10 days post-infection, the plants are uprooted, rinsed briefly and the roots frozen in liquid nitrogen. Approximately 20 grams of root tissue is harvested from the infected plants. The harvested tissue is then stored at -80° C. until RNA preparation.

[0482]The SOYMON030 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) flower bud tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. A total of 100 mg of flower buds are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0483]The SOYMON031 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) carpel and stamen tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. Flowers are dissected to separate petals, sepals and reproductive structures (carpels and stamens). A total of 300 mg of carpel and stamen tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0484]The SOYMON032 cDNA library is prepared from the Asgrow cultivar A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry soybean seed meristem tissue. Surface sterilized seeds are germinated in liquid media for 24 hours. The seed axis is then excised from the barely germinating seed, placed on tissue culture media and incubated overnight at 20° C. in the dark. The supportive tissue is removed from the explant prior to harvest. Approximately 570 mg of tissue is harvested and frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0485]The SOYMON033 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heat-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to an incubator set at 40° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at -80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance. Total RNA and poly A.sup.+ RNA is prepared from equal amounts of pooled tissue.

[0486]The SOYMON034 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) cold-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to a cold room set at 5° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at -80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance.

[0487]The SOYMON035 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed coat tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are harvested from mid to nearly full maturation (seed coats are not yellowing). The entire embryo proper is removed from the seed coat sample and the seed coat tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0488]The SOYMON036 cDNA library is generated from soybean cultivars PI171451, PI227687 and PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) insect challenged leaves. Plants from each of the three cultivars are grown in screenhouse conditions. The screenhouse is divided in half and one half of the screenhouse is infested with soybean looper and the other half infested with velvetbean caterpillar. A single leaf is taken from each of the representative plants at 3 different time points, 11 days after infestation, 2 weeks after infestation and 5 weeks after infestation and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation. Total RNA and poly A+ RNA is isolated from pooled tissue consisting of equal quantities of all 18 samples (3 genotypes×3 sample times×2 insect genotypes).

[0489]The SOYMON037 cDNA library is generated from soybean cultivar A3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) etiolated axis and radical tissue. Seeds are planted in moist vermiculite, wrapped and kept at room temperature in complete darkness until harvest. Etiolated axis and hypocotyl tissue is harvested at 2, 3 and 4 days post-planting. A total of 1 gram of each tissue type is harvested at 2, 3 and 4 days after planting and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0490]The SOYMON038 cDNA library is generated from soybean variety Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry seeds. Explants are prepared for transformation after germination of surface-sterilized seeds on solid tissue media. After 6 days, at 28° C. and 18 hours of light per day, the germinated seeds are cold shocked at 4° C. for 24 hours. Meristemic tissue and part of the hypocotyl is remove and cotyledon excised. The prepared explant is then wounded for Agrobacterium infection. The 2 grams of harvested tissue is frozen in liquid nitrogen and stored at -80° C. until RNA preparation.

[0491]The Soy51 (LIB3027) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

[0492]The Soy52 (LIB3028) cDNA library is generated from normalized flower DNA. Single stranded DNA representing approximately 1×10⁶ colony forming units of SOYMON022 harvested tissue is used as the starting material for normalization. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

[0493]The Soy53 (LIB3039) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling shoot apical meristem tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Apical tissue is harvested from seedling shoot meristem tissue, 7-8 days after the start of imbibition. The apex of each seedling is dissected to include the fifth node to the apical meristem. The fifth node corresponds to the third trifoliate leaf in the very early stages of development. Stipules completely envelop the leaf primordia at this time. A total of 200 mg of apical tissue is harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at -80° C. until RNA preparation.

[0494]The Soy54 (LIB3040) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heart to torpedo stage embryo tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected and embryos removed from surrounding endosperm and maternal tissues. Embryos from globular to young torpedo stages (by corresponding analogy to Arabidopsis) are collected with a bias towards the middle of this spectrum. Embryos which are beginning to show asymmetric development of cotyledons are considered the upper developmental boundary for the collection and are excluded. A total of 12 mg embryo tissue is frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0495]Soy55 (LIB3049) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) young seed tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected from very young pods (5 to 15 days after flowering). A total of 100 mg of seeds are harvested and frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0496]Soy56 (LIB3029) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are not converted to double stranded form and represent a non-normalized seed pool for comparison to Soy51 cDNA libraries.

[0497]The Soy58 (LIB3050) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed root tissue subtracted from control root tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days root tissue from both drought stressed and control (watered regularly) plants are collected and frozen in dry-ice. The harvested tissue is stored at -80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

[0498]The Soy59 (LIB3051) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) endosperm tissue. Seeds are germinated on paper towels under laboratory ambient light conditions. At 8, 10 and 14 hours after imbibition, the seed coats are harvested. The endosperm consists of a very thin layer of tissue affixed to the inside of the seed coat. The seed coat and endosperm are frozen immediately after harvest in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0499]The Soy60 (LIB3072) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed seed plus pod subtracted from control seed plus pod tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at -80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

[0500]The Soy61 (LIB3073) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 orjasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). For this library's construction, the eighth fraction of the cDNA size fractionation step was used for ligation.

[0501]The Soy62 (LIB3074) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 orjasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). For this library's construction, the ninth fraction of the cDNA size fractionation step was used for ligation.

[0502]The Soy65 (LIB3107) 07cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At the R3 stage of development, drought is imposed by withholding water. At 3, 4, 5 and 6 days, tissue is harvested and wilting is not obvious until the fourth day. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0503]The Soy66 (LIB3109) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) non-drought stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At 3, 4, 5 and 6 days, control abscission layer tissue is harvested. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0504]Soy67 (LIB3065) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

[0505]Soy68 (LIB3052) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

[0506]Soy69 (LIB3053) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) normalized leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4 h node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

[0507]Soy70 (LIB3055) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4 h node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0508]Soy71 (LIB3056) cDNA library is generated from soybean cultivars Cristalina and FT108 (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at -80° C. until RNA preparation.

[0509]Soy72 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed leaf control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at -80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

[0510]Soy73 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed leaf subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at -80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

[0511]The Soy76 (Lib3106) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid and arachidonic treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 orjasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.) in order to capture some of the smaller transcripts characteristic of antifungal proteins.

[0512]Soy77 (LIB3108) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)jasmonic acid control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0. 1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2× SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPORT vector in order to capture some of the smaller transcripts characteristic of antifungal proteins.

[0513]The Lib9 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, leaf tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Leaf blades were cut with sharp scissors at seven weeks after planting. The tissue was immediately frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

[0514]The Lib22 cDNA library is prepared from Arabidopsis thaliana Columbia ecotype, root tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. After 5-6 weeks the plants are in the reproductive growth phase. Stems are bolting from the base of the plants. After 7 weeks, more stems, floral buds appear, and a few flowers are starting to open. The 7-week old plants are rinsed intensively by tope water remove dirt from the roots, and blotted by paper towel. The tissues are immediately frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA preparation.

[0515]The Lib23 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, stem tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Stems were collected seven to eight weeks after planting by cutting the stems from the base and cutting the top of the plant to remove the floral tissue. The tissue was immediately frozen in liquid nitrogen and stored at -80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

[0516]The Lib24 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, flower bud tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Flower buds are green and unopened and harvested about seven weeks after planting. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

[0517]The Lib25 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, open flower tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Flowers are completely opened with all parts of floral structure observable, but no siliques are appearing. The tissue was immediately frozen in liquid nitrogen and stored at -80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

[0518]The Lib35 cDNA library of the present invention, was prepared from Arabidopsis thaliana Columbia ecotype leaf tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. After 5-6 weeks the plants are in the reproductive growth phase. Stems are bolting from the base of the plants. After 7 weeks, more stems and floral buds appeared and a few flowers were starting to open. Leaf blades were collected by cutting with sharp scissors. The tissues were immediately frozen in liquid nitrogen and stored at -80° C. until use. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

[0519]The Lib 146 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, immature seed tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. At approximately 7-8 weeks of age, the seeds are harvested. The seeds ranged in maturity from the smallest seeds that could be dissected from silques to just before starting to turn yellow in color. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is stored at -80° C. until RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library is normalized using a PCR-based protocol.

[0520]The Lib3032 (Lib80) cDNA libraries are generated from Brassica napus seeds harvested 30 days after pollination. The cDNA libraries are constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA is used as the starting material for cDNA synthesis, and first strand cDNA synthesis is carried out at 45° C.

[0521]The Lib3034 (Lib82) cDNA libraries are generated from Brassica napus seeds harvested 15 and 18 days after pollination. The cDNA libraries are constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA is used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

[0522]The Lib3099 cDNA library is generated by a subtraction procedure. The library contains cDNAs whose abundance is enriched in the Brassica napus 15 and 18 day after pollination seed tissues when compared to Brassica leaf tissues. The cDNA synthesis is performed on Brassica leaf RNA and Brassica RNA isolated from seeds harvested 15 and 18 days after pollination using a Smart PCR cDNA synthesis kit according to the manufacturers protocol (Clonetech, Palo Alto, Calif. U.S.A.). The subtracted cDNA is generated using the Clontech PCR-Select subtraction kit according to the manufacturers protocol (Clontech, Palo Alto, Calif. U.S.A.). The subtracted cDNA was cloned into plasmid vector pCR2.1 according to the manufacturers protocol (Invitrogen, Carlsbad, Calif. U.S.A.).

[0523]The Lib3033 (Lib81) cDNA libraries are generated from the Schizochytrium species cells. The Schizochytrium species cells are grown in liquid media until saturation. The culture is centrifuged to pellet the cells, the medium is decanted off, and pellet immediately frozen in liquid nitrogen. Wax esters are produced under such dark, anaerobic, rich-medium conditions. High wax production by the cultures is verified by microscopy (fluorescein staining of wax bodies) and by lipid extraction/TLC/GC. The harvested cells are stored at -80° C. until RNA preparation. RNA is prepared from the frozen Euglena cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

[0524]The Lib47 cDNA library is generated from Euglena gracilus strain 753 (ATTC No. 30285, ATCC Manassas, Va. U.S.A.) grown in liquid culture. A liquid culture is inoculated with 1/10 volume of a previously-grown saturated culture, and the new culture for 4 days under near-anaerobic conditions (near-anaerobic cultures are not agitated, just gently swirled once a day) in the dark in 2× Beef (10 g/l bacto peptone, 4 g/l yeast extract, 2 g/l beef extract, 6 g/l glucose). The culture is then centrifuged to pellet the cells, the medium is decanted off, and pellet immediately frozen in liquid nitrogen. Wax esters are produced under such dark, anaerobic, rich-medium conditions. High wax production by the cultures is verified by microscopy (fluorescein staining of wax bodies) and by lipid extraction/TLC/GC. The harvested cells are stored at -80° C. until RNA preparation. RNA is prepared from the frozen Euglena cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

[0525]The Lib44 cDNA library is generated from Phaeodactylum tricornatum grown in modified Jones medium for 3 days. The cells were harvested by centrifugation and the resulting pellet frozen immediately in liquid nitrogen. The harvested cells are stored at -80° C. until RNA preparation. RNA is prepared from the frozen Phaeodactylum cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45 degrees centigrade.

[0526]The LIB3036 genomic library is generated from Mycobacterium neoaurum US52 (ATCC No. 23072, ATCC, Manassas, Va. U.S.A.) cells. Mycobacterium neoaurum US52 is a gram-positive Actinomycete bacterium. Mycobacterium neoaurum US52 is genetically related to Mycobacterium tuberculosis, but there is no reason to believe that it is a primary pathogen. It normally is saprophytic, i.e. it lives in soil and gets nutrients from decaying matter. Genomic DNA obtained from Mycobacterium neoaurum US52 is digested for various times with the restriction enzyme Sau3A. The DNA fractions are size-separated on an agarose gel, and the first fraction wherein most of the partially-digested fragments are about 10 kB is used to isolated fragments in the range of 2-3 kB. For LIB3036, the 2-3 kB DNA is cloned into vector pRY401 (Invitrogen, Carlsbad, Calif. U.S.A.). The vector pZERO-2 (Invitrogen, Carlsbad, Calif. U.S.A.). is used for the construction of LIB3104.

[0527]The stored RNA is purified using Trizol reagent from Life Technologies (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.), essentially as recommended by the manufacturer. Poly A+ RNA (mRNA) is purified using magnetic oligo dT beads essentially as recommended by the manufacturer (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.).

[0528]Construction of plant cDNA libraries is well-known in the art and a number of cloning strategies exist. A number of cDNA library construction kits are commercially available. The Superscript® Plasmid System for cDNA synthesis and Plasmid Cloning (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.) is used, following the conditions suggested by the manufacturer.

[0529]Normalized libraries are made using essentially the Soares procedure (Soares et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:9228-9232 (1994), the entirety of which is herein incorporated by reference). This approach is designed to reduce the initial 10,000-fold variation in individual cDNA frequencies to achieve abundances within one order of magnitude while maintaining the overall sequence complexity of the library. In the normalization process, the prevalence of high-abundance cDNA clones decreases dramatically, clones with mid-level abundance are relatively unaffected and clones for rare transcripts are effectively increased in abundance.

Example 2

[0530]The cDNA libraries are plated on LB agar containing the appropriate antibiotics for selection and incubated at 37° for a sufficient time to allow the growth of individual colonies. Single colonies are individually placed in each well of a 96-well microtiter plates containing LB liquid including the selective antibiotics. The plates are incubated overnight at approximately 37° C. with gentle shaking to promote growth of the cultures. The plasmid DNA is isolated from each clone using Qiaprep plasmid isolation kits, using the conditions recommended by the manufacturer (Qiagen Inc., Santa Clara, Calif. U.S.A.).

[0531]Template plasmid DNA clones are used for subsequent sequencing. For sequencing, the ABI PRISM dRhodamine Terminator Cycle Sequencing Ready Reaction Kit with AmpliTaq® DNA Polymerase, FS, is used (PE Applied Biosystems, Foster City, Calif. U.S.A.).

Example 3

[0532]Nucleic acid sequences that encode for the following tetrapyrrole pathway enzymes: putative chlorophyll synthetase, protochlorophyllide reductase, putative protochlorophyllide reductase, coproporphyrinogen oxidase, protoporphyrinogen oxidase, uroporphyrinogen decarboxylase, putative uroporphyrinogen decarboxylase, porphobilinogen synthase enzyme, hydroxymethylbilane synthase enzyme, glutamate-1-semialdehyde 2,1-aminomutase enzyme, glutamate tRNA ligase enzyme, glutamyl-tRNA reductase enzyme, Mg-chelatase enzyme, and ferrochelatase enzyme are identified from the Monsanto EST PhytoSeq database using TBLASTN (default values)(TBLASTN compares a protein query against the six reading frames of a nucleic acid sequence). Matches found with BLAST P values equal or less than 0.001 (probability) or BLAST Score of equal or greater than 90 are classified as hits. If the program used to determine the hit is HMMSW then the score refers to HMMSW score.

[0533]In addition, the GenBank database is searched with BLASTN and BLASTX (default values) using ESTs as queries. EST that pass the hit probability threshold of 10e^-8 for the following enzymes are combined with the hits generated by using TBLASTN (described above) and classified by enzyme (see Table A below).

[0534]A cluster refers to a set of overlapping clones in the PhytoSeq database. Such an overlapping relationship among clones is designated as a "cluster" when BLAST scores from pairwise sequence comparisons of the member clones meets a predetermined minimum value or product score of 50 or more (Product Score=(BLAST SCORE×Percentage Identity)/(5×minimum [length (Seq1), length (Seq2)]))

[0535]Since clusters are formed on the basis of single-linkage relationships, it is possible for two non-overlapping clones to be members of the same cluster if, for instance, they both overlap a third clone with at least the predetermined minimum BLAST score (stringency). A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a "singleton"), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. Clones grouped in a cluster in most cases represent a contiguous sequence.

TABLE-US-00002 TABLE A* Seq No. Cluster ID Clone ID Library NCBI gi Method Score P-value % Ident SOYBEAN PUTATIVE CHLOROPHYLL SYNTHETASE 1 -700941050 700941050H1 SOYMON024 g972938 BLASTX 349 1e-41 75 2 -701212263 701212263H1 SOYMON035 g972938 BLASTX 75 1e-9 65 3 -701213734 701213734H1 SOYMON035 g972937 BLASTN 191 1e-27 83 4 14458 LIB3049-005- LIB3049 g3068709 BLASTX 101 1e-35 59 Q1-E1-F12 5 14458 700975706H1 SOYMON009 g972938 BLASTX 75 1e-9 50 6 14458 701047496H1 SOYMON032 g972938 BLASTX 75 1e-9 52 7 26375 701156709H1 SOYMON031 g972938 BLASTX 102 1e-15 92 8 26375 701156060H1 SOYMON031 g972937 BLASTN 275 1e-13 80 SOYBEAN PROTOCHLOROPHYLLIDE REDUCTASE 9 -700654876 700654876H1 SOYMON004 g20829 BLASTN 269 1e-23 82 10 -700657235 700657235H1 SOYMON004 g20829 BLASTN 728 1e-57 83 11 -700657437 700657437H1 SOYMON004 g20829 BLASTN 668 1e-46 84 12 -700757662 700757662H1 SOYMON015 g20829 BLASTN 1012 1e-83 89 13 -700842232 700842232H1 SOYMON020 g20829 BLASTN 442 1e-32 81 14 -700976426 700976426H1 SOYMON009 g2244613 BLASTN 1038 1e-77 85 15 11407 700652980H1 SOYMON003 g2244613 BLASTN 741 1e-52 71 16 11407 700735503H1 SOYMON010 g2244613 BLASTN 505 1e-33 70 17 11407 701142847H1 SOYMON038 g2244613 BLASTN 509 1e-33 67 18 11407 700652452H1 SOYMON003 g2244613 BLASTN 535 1e-33 69 19 11407 700735307H1 SOYMON010 g2244613 BLASTN 491 1e-32 70 20 11407 701107638H1 SOYMON036 g2244613 BLASTN 493 1e-32 70 21 11407 700952613H1 SOYMON022 g2244613 BLASTN 479 1e-31 70 22 11407 701118520H1 SOYMON037 g2244613 BLASTN 475 1e-29 70 23 11407 700731513H1 SOYMON010 g2244613 BLASTN 455 1e-28 72 24 11407 701037153H1 SOYMON029 g2244613 BLASTN 455 1e-27 72 25 11407 700838406H1 SOYMON020 g2244614 BLASTX 229 1e-24 57 26 11407 700736971H1 SOYMON010 g2244613 BLASTN 389 1e-22 75 27 11407 701208151H1 SOYMON035 g2244613 BLASTN 387 1e-21 76 28 11407 700658204H1 SOYMON004 g2244614 BLASTX 173 1e-16 61 29 11407 700657759H1 SOYMON004 g2244614 BLASTX 120 1e-14 54 30 11407 700854307H1 SOYMON023 g20829 BLASTN 190 1e-12 80 31 2160 LIB3039-002- LIB3039 g2244613 BLASTN 690 1e-46 84 Q1-E1-G10 32 2160 701107175H1 SOYMON036 g20829 BLASTN 628 1e-43 88 33 21731 700660488H1 SOYMON004 g20829 BLASTN 796 1e-59 86 34 21731 701134573H1 SOYMON038 g20829 BLASTN 646 1e-44 84 35 21739 700655688H1 SOYMON004 g2244613 BLASTN 298 1e-35 84 36 21739 700655588H1 SOYMON004 g20830 BLASTX 125 1e-21 88 37 2977 700763883H1 SOYMON018 g20829 BLASTN 892 1e-80 83 38 2977 701139350H1 SOYMON038 g20829 BLASTN 614 1e-72 88 39 2977 700849172H1 SOYMON021 g20829 BLASTN 956 1e-70 87 40 2977 700993334H1 SOYMON011 g20829 BLASTN 793 1e-69 87 41 2977 700980689H1 SOYMON009 g20829 BLASTN 765 1e-66 83 42 2977 700754834H1 SOYMON014 g20829 BLASTN 910 1e-66 86 43 2977 701054679H1 SOYMON032 g20829 BLASTN 501 1e-60 81 44 2977 701142549H1 SOYMON038 g20829 BLASTN 595 1e-59 82 45 2977 700909828H1 SOYMON022 g20829 BLASTN 695 1e-59 84 46 2977 701153047H1 SOYMON031 g20829 BLASTN 715 1e-50 87 47 2977 700981305H1 SOYMON009 g2244613 BLASTN 645 1e-44 70 48 2977 700737910H1 SOYMON012 g2244613 BLASTN 598 1e-41 69 49 2977 701106762H1 SOYMON036 g2244613 BLASTN 602 1e-41 70 50 2977 700893019H1 SOYMON024 g2244613 BLASTN 595 1e-40 70 51 2977 700888819H1 SOYMON024 g2244613 BLASTN 555 1e-37 69 52 2977 700557617H1 SOYMON001 g2244613 BLASTN 557 1e-37 69 53 2977 700989268H1 SOYMON011 g20829 BLASTN 297 1e-35 80 54 2977 700978858H1 SOYMON009 g2244613 BLASTN 529 1e-35 69 55 2977 701063251H1 SOYMON033 g2244613 BLASTN 525 1e-34 63 56 2977 700737989H1 SOYMON012 g20829 BLASTN 190 1e-33 72 57 2977 LIB3054-001- LIB3054 g2244613 BLASTN 487 1e-32 70 Q1-B1-A11 58 2977 701057704H1 SOYMON033 g2244613 BLASTN 470 1e-29 72 59 2977 701139740H1 SOYMON038 g2244613 BLASTN 477 1e-29 69 60 2977 LIB3039-043- LIB3039 g2244614 BLASTX 99 1e-28 55 Q1-E1-F3 61 2977 701105971H1 SOYMON036 g2244613 BLASTN 454 1e-28 71 62 2977 700789775H1 SOYMON011 g2244613 BLASTN 429 1e-26 72 63 2977 700732675H1 SOYMON010 g2244613 BLASTN 437 1e-26 69 64 2977 701137164H1 SOYMON038 g2244613 BLASTN 429 1e-25 72 65 2977 700788180H1 SOYMON011 g2244613 BLASTN 431 1e-25 72 66 2977 700680942H1 SOYMON008 g20829 BLASTN 349 1e-24 72 67 2977 700953017H1 SOYMON022 g2244613 BLASTN 395 1e-22 71 68 2977 700962368H1 SOYMON022 g2244613 BLASTN 395 1e-22 71 69 2977 700737258H1 SOYMON010 g2244613 BLASTN 395 1e-22 71 70 2977 701058308H1 SOYMON033 g2244613 BLASTN 244 1e-14 78 71 2977 701108820H1 SOYMON036 g968974 BLASTN 254 1e-14 76 72 2977 700658246H1 SOYMON004 g20830 BLASTX 123 1e-13 76 73 2977 700990646H1 SOYMON011 g20829 BLASTN 255 1e-12 92 74 2977 700548092H1 SOYMON001 g20830 BLASTX 92 1e-11 79 75 2977 701136902H1 SOYMON038 g2244613 BLASTN 265 1e-11 69 76 2977 701152877H1 SOYMON031 g20830 BLASTX 128 1e-10 76 77 2977 700994862H1 SOYMON011 g20830 BLASTX 128 1e-10 76 78 2977 701148824H1 SOYMON031 g20830 BLASTX 128 1e-10 76 79 2977 701047440H1 SOYMON032 g20830 BLASTX 128 1e-10 76 80 2977 700556683H1 SOYMON001 g968974 BLASTN 252 1e-10 76 81 2977 701146931H1 SOYMON031 g2244613 BLASTN 253 1e-10 72 82 2977 701142178H1 SOYMON038 g20830 BLASTX 122 1e-9 73 83 2977 701152593H1 SOYMON031 g20830 BLASTX 123 1e-9 75 84 2977 700737725H1 SOYMON012 g20829 BLASTN 218 1e-9 74 85 2977 700683007H1 SOYMON008 g2244613 BLASTN 241 1e-9 70 86 2977 700739072H1 SOYMON012 g2244613 BLASTN 244 1e-9 71 87 4903 700658027H1 SOYMON004 g20829 BLASTN 820 1e-59 79 88 4903 700852934H1 SOYMON023 g20829 BLASTN 453 1e-48 78 89 6970 LIB3052-012- LIB3052 g968974 BLASTN 934 1e-69 78 Q1-N1-A11 90 6970 700660679H1 SOYMON004 g20829 BLASTN 862 1e-67 87 91 6970 700682420H2 SOYMON008 g968976 BLASTN 864 1e-63 80 92 6970 700979758H2 SOYMON009 g2244613 BLASTN 865 1e-63 82 93 6970 700790842H1 SOYMON011 g968974 BLASTN 642 1e-57 82 94 6970 700994812H1 SOYMON011 g968976 BLASTN 423 1e-43 78 SOYBEAN PUTATIVE PROTOCHLOROPHYLLIDE REDUCTASE 95 -701065431 701065431H1 SOYMON034 g348719 BLASTN 767 1e-55 83 96 4640 700982771H1 SOYMON009 g348718 BLASTX 162 1e-15 93 SOYBEAN COPROPORPHYRINOGEN OXIDASE 97 -700671956 700671956H1 SOYMON006 g414665 BLASTN 291 1e-16 96 98 -701053612 701053612H1 SOYMON032 g414665 BLASTN 335 1e-27 94 99 -701208513 701208513H1 SOYMON035 g414665 BLASTN 639 1e-92 94 100 11665 700656318H1 SOYMON004 g414665 BLASTN 656 1e-93 98 101 11665 700964466H1 SOYMON022 g414665 BLASTN 611 1e-88 98 102 11665 700899782H1 SOYMON027 g414665 BLASTN 648 1e-87 98 103 11665 700844365H1 SOYMON021 g414665 BLASTN 648 1e-83 98 104 11665 701146220H1 SOYMON031 g414665 BLASTN 630 1e-75 98 105 11665 700660179H1 SOYMON004 g414665 BLASTN 530 1e-55 94 106 11665 700662658H1 SOYMON005 g1213066 BLASTN 742 1e-53 78 107 11665 701152413H1 SOYMON031 g1213066 BLASTN 742 1e-53 78 108 6121 LIB3065-002- LIB3065 g414665 BLASTN 1383 1e-128 96 Q1-N1-G8 109 6121 701108945H1 SOYMON036 g414665 BLASTN 1388 1e-106 98 110 6121 700789601H2 SOYMON011 g414665 BLASTN 1301 1e-99 99 111 6121 700994436H1 SOYMON011 g414665 BLASTN 858 1e-97 98 112 6121 700747416H1 SOYMON013 g414665 BLASTN 925 1e-94 100 113 6121 700978804H1 SOYMON009 g414665 BLASTN 941 1e-89 95 114 6121 701109318H1 SOYMON036 g414665 BLASTN 729 1e-85 96 115 6121 700873742H1 SOYMON018 g414665 BLASTN 484 1e-82 94 116 6121 701209226H1 SOYMON035 g414665 BLASTN 789 1e-81 97 117 6121 701060931H1 SOYMON033 g414665 BLASTN 768 1e-79 96 118 6121 701066887H1 SOYMON034 g414665 BLASTN 262 1e-68 90 119 6121 700899224H1 SOYMON027 g414665 BLASTN 434 1e-57 84 120 6121 700906273H1 SOYMON022 g414665 BLASTN 555 1e-37 97 121 6121 700992008H1 SOYMON011 g414665 BLASTN 560 1e-37 97 122 6121 700786848H2 SOYMON011 g414665 BLASTN 408 1e-25 98 123 6121 700734585H1 SOYMON010 g414665 BLASTN 250 1e-16 100 124 7272 700786276H2 SOYMON011 g414665 BLASTN 1170 1e-88 97 125 7272 700683451H1 SOYMON008 g414665 BLASTN 1019 1e-87 97 126 7272 700662424H1 SOYMON005 g414665 BLASTN 932 1e-84 98 127 7882 700680869H1 SOYMON008 g414665 BLASTN 763 1e-54 99 128 7882 700680628H1 SOYMON008 g414665 BLASTN 516 1e-34 94 SOYBEAN PROTOPORPHYRINOGEN OXIDASE 129 -700657957 700657957H1 SOYMON004 g1183006 BLASTN 729 1e-51 76 130 -700681258 700681258H1 SOYMON008 g1183006 BLASTN 651 1e-46 76 131 -701063830 701063830H1 SOYMON034 g2370335 BLASTX 142 1e-15 79 SOYBEAN UROPORPHYRINOGEN DECARBOXYLASE 132 -700730557 700730557H1 SOYMON009 g1009428 BLASTN 444 1e-38 69 133 -700789740 700789740H1 SOYMON011 g1009428 BLASTN 760 1e-54 80 134 -700974704 700974704H1 SOYMON005 g1016347 BLASTX 272 1e-30 55 135 -701048641 701048641H1 SOYMON032 g1009427 BLASTN 580 1e-39 71 136 -GM17920 LIB3055-003- LIB3055 g142136 BLASTX 97 1e-29 61 Q1-N1-H10 137 19517 701104233H1 SOYMON036 g1009429 BLASTX 228 1e-24 49 138 19517 701000103H1 SOYMON018 g1009429 BLASTX 167 1e-22 46 139 19517 701108875H1 SOYMON036 g1009429 BLASTX 137 1e-19 49 140 19517 700737952H1 SOYMON012 g1009429 BLASTX 188 1e-18 39 141 4729 700753974H1 SOYMON014 g1009427 BLASTN 816 1e-59 82 142 4729 701126044H1 SOYMON037 g1009427 BLASTN 799 1e-57 82 143 4729 700870535H1 SOYMON018 g1009427 BLASTN 405 1e-54 82 144 8117 700752125H1 SOYMON014 g1009428 BLASTN 444 1e-26 75 SOYBEAN PORPHOBILINOGEN SYNTHASE 145 -700678901 700678901H1 SOYMON007 g493019 BLASTN 1259 1e-101 97 146 -700680455 700680455H1 SOYMON008 g493019 BLASTN 915 1e-105 98 147 -700897467 700897467H1 SOYMON027 g493019 BLASTN 1091 1e-98 97 148 -700994415 700994415H1 SOYMON011 g493019 BLASTN 381 1e-21 97 149 -701002563 701002563H1 SOYMON018 g493019 BLASTN 608 1e-41 96 150 -701208590 701208590H1 SOYMON035 g493019 BLASTN 366 1e-21 92 151 -GM8017 LIB3039-038- LIB3039 g493019 BLASTN 224 1e-29 86 Q1-E1-H8 152 -GM9259 LIB3049-002- LIB3049 g493019 BLASTN 281 1e-16 84 Q1-E1-G5 153 -GM9536 LIB3049-003- LIB3049 g493019 BLASTN 426 1e-61 82 Q1-E1-D4 154 11129 700660017H1 SOYMON004 g313724 BLASTX 176 1e-17 51 155 22115 701208693H1 SOYMON035 g493019 BLASTN 1353 1e-103 97 156 22115 701151960H1 SOYMON031 g493019 BLASTN 1245 1e-94 100 157 22115 700846243H1 SOYMON021 g493019 BLASTN 571 1e-75 99 158 23112 701207084H1 SOYMON035 g493019 BLASTN 1408 1e-108 97 159 23112 700654971H1 SOYMON004 g493019 BLASTN 1236 1e-94 98 160 25460 700656593H1 SOYMON004 g493019 BLASTN 1296 1e-99 99 161 25460 701050212H1 SOYMON032 g493019 BLASTN 1235 1e-94 98 162 25460 701123120H1 SOYMON037 g493019 BLASTN 1245 1e-94 100 163 25460 701055012H1 SOYMON032 g493019 BLASTN 868 1e-93 99 164 3678 LIB3039-036- LIB3039 g493019 BLASTN 1757 1e-137 99 Q1-E1-D2 165 3678 LIB3039-031- LIB3039 g493019 BLASTN 1736 1e-135 99 Q1-E1-F9 166 3678 700553643H1 SOYMON001 g493019 BLASTN 1383 1e-106 98 167 3678 700558620H1 SOYMON001 g493019 BLASTN 1361 1e-104 98 168 3678 701046832H1 SOYMON032 g493019 BLASTN 1185 1e-100 96 169 3678 701109455H1 SOYMON036 g493019 BLASTN 1282 1e-98 97 170 3678 LIB3056-002- LIB3056 g493019 BLASTN 1286 1e-98 98 Q1-B1-D5 171 3678 700844432H1 SOYMON021 g493019 BLASTN 1274 1e-97 99 172 3678 700847337H1 SOYMON021 g493019 BLASTN 1242 1e-94 98 173 3678 700994748H1 SOYMON011 g493019 BLASTN 1221 1e-92 98 174 3678 701213656H1 SOYMON035 g493019 BLASTN 1176 1e-89 99 175 3678 700969539H1 SOYMON005 g493019 BLASTN 1120 1e-88 95 176 3678 700862858H1 SOYMON020 g493019 BLASTN 775 1e-85 98 177 3678 701109689H1 SOYMON036 g493019 BLASTN 968 1e-85 97 178 3678 701105613H1 SOYMON036 g493019 BLASTN 1127 1e-85 98 179 3678 700762772H1 SOYMON015 g493019 BLASTN 1039 1e-84 97 180 3678 LIB3039-047- LIB3039 g493019 BLASTN 619 1e-82 94 Q1-E1-D4 181 3678 700962419H1 SOYMON022 g493019 BLASTN 670 1e-82 99 182 3678 700975590H1 SOYMON009 g493019 BLASTN 965 1e-82 93 183 3678 701108204H1 SOYMON036 g493019 BLASTN 1069 1e-80 99 184 3678 700725416H1 SOYMON009 g493019 BLASTN 633 1e-66 89 185 3678 701055787H1 SOYMON032 g493019 BLASTN 790 1e-57 100 186 3678 700996808H1 SOYMON018 g493019 BLASTN 740 1e-52 100 187 3678 701210127H1 SOYMON035 g493019 BLASTN 545 1e-36 100 188 3678 700739360H1 SOYMON012 g493019 BLASTN 245 1e-21 99 189 3678 700742044H1 SOYMON012 g493019 BLASTN 313 1e-17 98 190 3678 701065288H1 SOYMON034 g493019 BLASTN 335 1e-17 100 191 3678 701110261H1 SOYMON036 g493019 BLASTN 211 1e-12 99 SOYBEAN HYDROXYMETHYLBILANE SYNTHASE 154 11129 700660017H1 SOYMON004 g313724 BLASTX 176 1e-17 51 192 -700653648 700653648H1 SOYMON003 g313723 BLASTN 262 1e-60 90 193 -700666293 700666293H1 SOYMON005 g313723 BLASTN 772 1e-65 82 194 -700975534 700975534H1 SOYMON009 g313723 BLASTN 498 1e-76 89 195 -701064190 701064190H1 SOYMON034 g313723 BLASTN 450 1e-62 85 196 -701125566 701125566H1 SOYMON037 g313723 BLASTN 474 1e-55 83 197 11129 700656680H1 SOYMON004 g313723 BLASTN 834 1e-73 85 198 12006 700556506H1 SOYMON001 g313723 BLASTN 652 1e-69 83 199 12006 700557751H1 SOYMON001 g313723 BLASTN 652 1e-65 83 200 12006 700556513H1 SOYMON001 g313723 BLASTN 572 1e-61 83 201 12006 700848102H1 SOYMON021 g313723 BLASTN 459 1e-54 81 202 20966 701108946H1 SOYMON036 g313150 BLASTX 148 1e-13 83 203 20966 701054125H1 SOYMON032 g313150 BLASTX 148 1e-13 83 204 20966 701108239H1 SOYMON036 g313150 BLASTX 122 1e-9 83 205 8428 LIB3052-015- LIB3052 g313723 BLASTN 865 1e-79 79 Q1-N1-G5 206 8428 LIB3055-013- LIB3055 g313723 BLASTN 1021 1e-76 80 Q1-N1-H6 207 8428 701140841H1 SOYMON038 g313723 BLASTN 942 1e-69 85 208 8428 700559220H1 SOYMON001 g313723 BLASTN 871 1e-63 82 209 8428 700998668H1 SOYMON018 g313723 BLASTN 872 1e-63 84 210 8428 701047766H1 SOYMON032 g313723 BLASTN 855 1e-62 85 211 8428 701055336H1 SOYMON032 g313723 BLASTN 848 1e-61 85 212 8428 700558405H1 SOYMON001 g313723 BLASTN 733 1e-57 85 213 8428 700758672H1 SOYMON015 g313723 BLASTN 406 1e-50 85 214 8428 700904365H1 SOYMON022 g313723 BLASTN 392 1e-48 85 215 8428 700987727H1 SOYMON009 g313723 BLASTN 563 1e-38 84 216 8428 701119125H1 SOYMON037 g313723 BLASTN 386 1e-21 84 217 8428 700833610H1 SOYMON019 g313723 BLASTN 314 1e-17 84 SOYBEAN GLUTAMATE-1-SEMIALDEHYDE 2,1-AMINOMUTASE 218 -700659530 700659530H1 SOYMON004 g310566 BLASTN 1138 1e-86 94 219 -700733492 700733492H1 SOYMON010 g310566 BLASTN 552 1e-47 87

220 -700737926 700737926H1 SOYMON012 g310566 BLASTN 421 1e-34 84 221 -700757107 700757107H1 SOYMON015 g310566 BLASTN 640 1e-91 96 222 -700852479 700852479H1 SOYMON023 g310566 BLASTN 1162 1e-88 98 223 -700971066 700971066H1 SOYMON005 g310566 BLASTN 1208 1e-91 96 224 -700982010 700982010H1 SOYMON009 g310566 BLASTN 1016 1e-90 97 225 -700986986 700986986H1 SOYMON009 g310566 BLASTN 1313 1e-100 95 226 -701042351 701042351H1 SOYMON029 g310566 BLASTN 1238 1e-94 99 227 10046 LIB3051-102- LIB3051 g747967 BLASTN 549 1e-106 90 Q1-K1-E2 228 10046 700554665H1 SOYMON001 g310566 BLASTN 981 1e-86 94 229 10046 LIB3040-061- LIB3040 g310566 BLASTN 441 1e-80 83 Q1-E11-A4 230 10046 700560249H1 SOYMON001 g310566 BLASTN 773 1e-70 91 231 10046 700995521H1 SOYMON011 g310566 BLASTN 773 1e-70 91 232 10046 700741513H1 SOYMON012 g310566 BLASTN 608 1e-69 90 233 10046 701207555H1 SOYMON035 g310566 BLASTN 748 1e-68 91 234 10046 701109782H1 SOYMON036 g310566 BLASTN 615 1e-67 90 235 10046 701047870H1 SOYMON032 g310566 BLASTN 493 1e-57 89 236 10046 701108348H1 SOYMON036 g310566 BLASTN 441 1e-51 88 237 10046 701041675H1 SOYMON029 g310566 BLASTN 613 1e-51 89 238 10046 700659465H1 SOYMON004 g310566 BLASTN 407 1e-30 91 239 10046 701144580H1 SOYMON031 g310566 BLASTN 218 1e-9 89 240 11600 700788311H1 SOYMON011 g310566 BLASTN 1234 1e-94 95 241 11600 700902195H1 SOYMON027 g310566 BLASTN 1209 1e-91 96 242 11600 701135233H1 SOYMON038 g310566 BLASTN 1177 1e-89 96 243 12473 701104293H1 SOYMON036 g310566 BLASTN 884 1e-83 93 244 12473 701104392H1 SOYMON036 g310566 BLASTN 754 1e-76 93 245 13619 700877188H1 SOYMON018 g310566 BLASTN 1323 1e-101 99 246 13619 700845619H1 SOYMON021 g310566 BLASTN 948 1e-70 93 247 20047 700660491H1 SOYMON004 g310566 BLASTN 718 1e-67 93 248 20047 700989453H1 SOYMON011 g310566 BLASTN 369 1e-21 95 249 5811 LIB3049-032- LIB3049 g747967 BLASTN 1188 1e-131 96 Q1-E1-A12 250 5811 LIB3049-030- LIB3049 g747967 BLASTN 1245 1e-98 100 Q1-E1-C2 251 5811 701142371H1 SOYMON038 g747967 BLASTN 1209 1e-93 96 252 5811 701155760H1 SOYMON031 g747967 BLASTN 1215 1e-92 100 253 5811 700983730H1 SOYMON009 g310566 BLASTN 417 1e-85 96 254 5811 701064792H1 SOYMON034 g310566 BLASTN 952 1e-81 97 255 5811 700561468H1 SOYMON002 g747967 BLASTN 757 1e-75 90 256 5811 700945376H1 SOYMON024 g310566 BLASTN 884 1e-73 98 257 5811 700756262H1 SOYMON014 g747967 BLASTN 722 1e-72 88 258 5811 700981893H1 SOYMON009 g747967 BLASTN 610 1e-70 92 259 5811 700562668H1 SOYMON002 g747967 BLASTN 807 1e-58 98 260 5811 700979322H1 SOYMON009 g747967 BLASTN 465 1e-54 97 261 5811 700905434H1 SOYMON022 g747967 BLASTN 707 1e-50 97 262 5811 700733377H1 SOYMON010 g747967 BLASTN 640 1e-48 93 263 5811 700562340H1 SOYMON002 g747967 BLASTN 629 1e-43 97 264 5811 701211223H1 SOYMON035 g310566 BLASTN 506 1e-41 94 265 5811 700565140H1 SOYMON002 g310566 BLASTN 416 1e-34 91 SOYBEAN GLUTAMATE tRNA LIGASE 266 -700653562 700653562H1 SOYMON003 g1008482 BLASTN 280 1e-27 64 267 -700740810 700740810H1 SOYMON012 g2370487 BLASTX 217 1e-22 43 268 -700893754 700893754H1 SOYMON024 g1008483 BLASTX 121 1e-8 34 269 -701009959 701009959H2 SOYMON019 g603849 BLASTX 175 1e-17 97 270 -701011820 701011820H1 SOYMON019 g1322915 BLASTX 129 1e-20 58 271 -701051674 701051674H1 SOYMON032 g2370487 BLASTX 221 1e-23 57 272 -701052937 701052937H1 SOYMON032 g1322915 BLASTX 236 1e-30 69 273 -701060112 701060112H1 SOYMON033 g1322915 BLASTX 121 1e-20 48 274 -701109025 701109025H1 SOYMON036 g157564 BLASTX 179 1e-18 67 275 -GM18124 LIB3065-002- LIB3065 g2995454 BLASTN 758 1e-66 78 Q1-N1-C2 276 -GM36590 LIB3051-050- LIB3051 g603849 BLASTX 91 1e-37 67 Q1-K1-D3 277 20438 700976589H1 SOYMON009 g2370487 BLASTX 253 1e-31 59 278 24353 701054537H1 SOYMON032 g157564 BLASTX 232 1e-24 56 279 24353 701054523H1 SOYMON032 g157564 BLASTX 192 1e-19 70 280 24353 701054530H1 SOYMON032 g157564 BLASTX 195 1e-19 69 281 27156 701137188H1 SOYMON038 g1008483 BLASTX 246 1e-26 53 282 27156 701207661H1 SOYMON035 g1008483 BLASTX 211 1e-24 52 283 27156 700726789H1 SOYMON009 g1008483 BLASTX 182 1e-22 52 284 32173 701202848H1 SOYMON035 g416260 BLASTN 443 1e-54 78 285 32173 LIB3049-020- LIB3049 g416260 BLASTN 733 1e-52 77 Q1-E1-G4 286 32173 700846868H1 SOYMON021 g157564 BLASTX 136 1e-20 67 287 7264 LIB3051-061- LIB3051 g2995454 BLASTN 841 1e-100 81 Q1-K1-B9 288 7712 700666928H1 SOYMON005 g157564 BLASTX 276 1e-30 56 289 7712 700665965H1 SOYMON005 g157564 BLASTX 263 1e-29 55 SOYBEAN GLUTAMYL-tRNA REDUCTASE 290 -700670004 700670004H1 SOYMON006 g1694925 BLASTN 397 1e-34 75 291 -700728413 700728413H1 SOYMON009 g1049056 BLASTN 696 1e-49 80 292 -700994105 700994105H1 SOYMON011 g1694925 BLASTN 771 1e-55 82 293 -700995896 700995896H1 SOYMON011 g1049057 BLASTX 84 1e-13 86 294 -701099031 701099031H1 SOYMON028 g1015318 BLASTN 459 1e-28 69 295 -701128557 701128557H1 SOYMON037 g1694925 BLASTN 432 1e-27 79 296 -GM35481 LIB3051-036- LIB3051 g1694925 BLASTN 1070 1e-80 73 Q1-K1-G6 297 25545 701123925H1 SOYMON037 g1694925 BLASTN 719 1e-51 78 298 25545 700727539H1 SOYMON009 g1694925 BLASTN 639 1e-44 79 299 2655 700553888H1 SOYMON001 g1694925 BLASTN 242 1e-20 74 300 2655 700553887H1 SOYMON001 g1694926 BLASTX 74 1e-8 93 301 2885 700728635H1 SOYMON009 g1015318 BLASTN 816 1e-59 78 302 2885 701097007H1 SOYMON028 g1015318 BLASTN 706 1e-50 80 303 3203 700986371H1 SOYMON009 g1694925 BLASTN 501 1e-69 80 304 3203 700556832H1 SOYMON001 g1694925 BLASTN 446 1e-47 81 305 3203 700995693H1 SOYMON011 g1039331 BLASTN 390 1e-39 72 306 33811 LIB3051-105- LIB3051 g1694925 BLASTN 573 1e-36 78 Q1-K1-B10 SOYBEAN Mg-CHELATASE 307 -700554488 700554488H1 SOYMON001 g1732468 BLASTN 835 1e-66 85 308 -700657604 700657604H1 SOYMON004 g2318116 BLASTN 975 1e-72 89 309 -700658239 700658239H1 SOYMON004 g1732468 BLASTN 1095 1e-92 100 310 -700737719 700737719H1 SOYMON012 g2318116 BLASTN 973 1e-72 87 311 -700902101 700902101H1 SOYMON027 g1732468 BLASTN 181 1e-15 89 312 -700943788 700943788H1 SOYMON024 g1732468 BLASTN 402 1e-23 91 313 -700992328 700992328H1 SOYMON011 g1732468 BLASTN 856 1e-62 87 314 -700996107 700996107H1 SOYMON018 g1732468 BLASTN 1208 1e-96 95 315 -701050458 701050458H1 SOYMON032 g1732468 BLASTN 881 1e-67 97 316 -701053810 701053810H1 SOYMON032 g1732468 BLASTN 1133 1e-99 99 317 -701119309 701119309H1 SOYMON037 g2318116 BLASTN 944 1e-83 91 318 -701128728 701128728H1 SOYMON037 g1732468 BLASTN 358 1e-28 90 319 -701134728 701134728H2 SOYMON038 g1732468 BLASTN 994 1e-85 95 320 -GM16990 LIB3055-002- LIB3055 g3059094 BLASTN 1141 1e-86 83 Q1-B1-F10 321 -GM18022 LIB3055-011- LIB3055 g3059094 BLASTN 1079 1e-138 99 Q1-N1-A4 322 -GM1974 LIB3028-009- LIB3028 g3059094 BLASTN 561 1e-66 89 Q1-B1-G12 323 -GM22404 LIB3030-009- LIB3030 g3059094 BLASTN 1104 1e-93 83 Q1-B1-A1 324 20034 700944453H1 SOYMON024 g2318116 BLASTN 1089 1e-81 90 325 20034 700834945H1 SOYMON019 g2318116 BLASTN 944 1e-69 87 326 20915 700993045H1 SOYMON011 g1732468 BLASTN 412 1e-23 97 327 20915 700978188H1 SOYMON009 g1732468 BLASTN 400 1e-22 95 328 22044 LIB3028-007- LIB3028 g2318116 BLASTN 1719 1e-134 89 Q1-B1-A8 329 22044 701063842H1 SOYMON034 g2318116 BLASTN 1084 1e-81 92 330 22044 700725383H1 SOYMON009 g2318116 BLASTN 1028 1e-76 90 331 22044 701001558H1 SOYMON018 g2318116 BLASTN 668 1e-75 89 332 26346 LIB3039-010- LIB3039 g1732468 BLASTN 441 1e-25 84 Q1-E1-G5 333 26346 700972265H1 SOYMON005 g1732468 BLASTN 287 1e-12 84 334 26346 701052502H1 SOYMON032 g1732468 BLASTN 241 1e-9 82 335 2822 LIB3054-010- LIB3054 g1732468 BLASTN 850 1e-120 93 Q1-N1-G10 336 2822 LIB3065-011- LIB3065 g1732468 BLASTN 841 1e-104 83 Q1-N1-D8 337 2822 700686645H1 SOYMON008 g1732468 BLASTN 868 1e-82 91 338 2822 700997745H1 SOYMON018 g1732468 BLASTN 878 1e-81 92 339 2822 700994426H1 SOYMON011 g1732468 BLASTN 1084 1e-81 89 340 2822 700739971H1 SOYMON012 g1732468 BLASTN 871 1e-80 95 341 2822 701138070H1 SOYMON038 g1732468 BLASTN 449 1e-65 87 342 2822 701203251H1 SOYMON035 g1732468 BLASTN 605 1e-65 87 343 2822 701206105H1 SOYMON035 g1732468 BLASTN 698 1e-65 88 344 2822 700895378H1 SOYMON027 g1732468 BLASTN 486 1e-62 83 345 2822 701105638H1 SOYMON036 g1732468 BLASTN 739 1e-61 90 346 2822 700898271H1 SOYMON027 g1732468 BLASTN 746 1e-61 89 347 2822 LIB3040-024- LIB3040 g1732468 BLASTN 652 1e-49 84 Q1-E1-H2 348 2822 700898124H1 SOYMON027 g1732468 BLASTN 578 1e-39 80 349 2822 700901555H1 SOYMON027 g1732468 BLASTN 529 1e-35 97 350 2822 700743195H1 SOYMON012 g1732468 BLASTN 324 1e-33 90 351 2822 700740845H1 SOYMON012 g1732468 BLASTN 478 1e-29 97 352 2822 700995694H1 SOYMON011 g1732468 BLASTN 314 1e-27 90 353 2822 700760635H1 SOYMON015 g1732468 BLASTN 438 1e-27 88 354 2822 701208243H1 SOYMON035 g1732468 BLASTN 429 1e-26 90 355 2822 700992554H1 SOYMON011 g1732468 BLASTN 348 1e-20 89 356 33722 LIB3030-005- LIB3030 g2318116 BLASTN 620 1e-48 81 Q1-B1-F12 357 33722 700653412H1 SOYMON003 g2318116 BLASTN 514 1e-32 88 358 4037 700982624H1 SOYMON009 g1732468 BLASTN 1353 1e-103 96 359 4037 701136660H1 SOYMON038 g1732468 BLASTN 1316 1e-100 96 360 4037 700979310H1 SOYMON009 g1732468 BLASTN 1010 1e-98 99 361 4037 700978952H1 SOYMON009 g1732468 BLASTN 1016 1e-91 92 362 4037 701104668H1 SOYMON036 g1732468 BLASTN 1152 1e-87 93 363 4037 700557049H1 SOYMON001 g1732468 BLASTN 1060 1e-79 92 364 4037 701107647H1 SOYMON036 g1732468 BLASTN 1030 1e-77 93 365 4037 701150771H1 SOYMON031 g1732468 BLASTN 1005 1e-74 92 366 4037 701154966H1 SOYMON031 g1732468 BLASTN 985 1e-73 100 367 4037 700989839H1 SOYMON011 g1732468 BLASTN 736 1e-52 93 368 4037 700756564H1 SOYMON014 g1732468 BLASTN 609 1e-41 93 369 4037 700753388H1 SOYMON014 g1732468 BLASTN 563 1e-38 93 370 4037 700850857H1 SOYMON023 g1732468 BLASTN 493 1e-32 92 371 4037 701150639H1 SOYMON031 g1732468 BLASTN 148 1e-17 82 SOYBEAN FERROCHELATASE 372 -700839666 700839666H1 SOYMON020 g2623989 BLASTN 736 1e-52 82 373 -700846363 700846363H1 SOYMON021 g439482 BLASTN 732 1e-52 77 374 -700901570 700901570H1 SOYMON027 g2623989 BLASTN 848 1e-61 81 375 -700907558 700907558H1 SOYMON022 g439482 BLASTN 700 1e-49 75 376 -701048026 701048026H1 SOYMON032 g439482 BLASTN 654 1e-45 71 377 -701064702 701064702H1 SOYMON034 g439482 BLASTN 439 1e-26 68 378 -701105159 701105159H1 SOYMON036 g2429617 BLASTN 487 1e-50 77 379 26592 701208376H1 SOYMON035 g439482 BLASTN 722 1e-51 78 380 26592 701097475H1 SOYMON028 g439482 BLASTN 729 1e-51 75 381 26592 701119601H1 SOYMON037 g439482 BLASTN 518 1e-40 78 382 28079 701015447H1 SOYMON019 g439482 BLASTN 840 1e-61 81 383 28079 701102766H1 SOYMON028 g439482 BLASTN 789 1e-56 82 MAIZE PUTATIVE CHLOROPHYLL SYNTHETASE 384 -700214815 700214815H1 SATMON016 g972938 BLASTX 244 1e-26 80 385 -700222875 700222875H1 SATMON011 g972937 BLASTN 340 1e-27 77 386 -L30662921 LIB3066-008- LIB3066 g3068702 BLASTN 504 1e-31 69 Q1-K1-C6 387 11381 LIB3078-007- LIB3078 g3068702 BLASTN 281 1e-57 75 Q1-K1-H2 388 11381 700084837H1 SATMON011 g972937 BLASTN 281 1e-48 74 389 11381 700088129H1 SATMON011 g972937 BLASTN 317 1e-44 78 390 11381 700045204H1 SATMON004 g972938 BLASTX 363 1e-43 73 391 11381 700084253H1 SATMON011 g972937 BLASTN 317 1e-35 79 392 11381 700427169H1 SATMONN01 g972938 BLASTX 225 1e-33 73 393 11381 700104418H1 SATMON010 g972937 BLASTN 317 1e-26 77 394 17510 700218357H1 SATMON016 g972937 BLASTN 173 1e-12 72 395 17510 700217457H1 SATMON016 g972937 BLASTN 173 1e-11 72 396 1913 700243564H1 SATMON010 g972937 BLASTN 357 1e-38 78 397 1913 700577332H1 SATMON031 g972938 BLASTX 165 1e-15 68 MAIZE PROTOCHLOROPHYLLIDE REDUCTASE 398 -700346250 700346250H1 SATMON021 g16117 BLASTN 338 1e-47 86 399 -700430255 700430255H1 SATMONN01 g19060 BLASTN 619 1e-42 93 400 -L30681828 LIB3068-021- LIB3068 g19060 BLASTN 212 1e-14 69 Q1-K1-A7 401 -L30782362 LIB3078-006- LIB3078 g19060 BLASTN 472 1e-47 73 Q1-K1-G6 402 18503 700321674H1 SATMON025 g683475 BLASTN 260 1e-50 82 403 18503 700220580H1 SATMON011 g683475 BLASTN 179 1e-16 80 404 2096 LIB36-005-Q1- LIB36 g683475 BLASTN 1633 1e-127 90 E1-G5 405 2096 LIB3062-018- LIB3062 g510676 BLASTN 1436 1e-116 84 Q1-K1-E9 406 2096 LIB3078-008- LIB3078 g19060 BLASTN 1017 1e-115 85 Q1-K1-H8 407 2096 LIB3078-004- LIB3078 g19060 BLASTN 1325 1e-101 84 Q1-K1-D7 408 2096 LIB3062-010- LIB3062 g19060 BLASTN 1159 1e-90 81 Q1-K1-G7 409 2096 700043426H1 SATMON004 g683475 BLASTN 1189 1e-90 93 410 2096 700093887H1 SATMON008 g19060 BLASTN 1194 1e-90 87 411 2096 700045326H1 SATMON004 g19060 BLASTN 1115 1e-84 92 412 2096 700439164H1 SATMON026 g683475 BLASTN 620 1e-81 89 413 2096 700093546H1 SATMON008 g19060 BLASTN 821 1e-81 88 414 2096 700081986H1 SATMON011 g510676 BLASTN 1076 1e-80 85 415 2096 700044764H1 SATMON004 g683475 BLASTN 1056 1e-79 89 416 2096 700098537H1 SATMON009 g510676 BLASTN 760 1e-73 87 417 2096 700340891H1 SATMON020 g683475 BLASTN 591 1e-70 88 418 2096 700100677H1 SATMON009 g683475 BLASTN 890 1e-65 89 419 2096 700265286H1 SATMON017 g683475 BLASTN 578 1e-53 88 420 2096 700212436H1 SATMON016 g16117 BLASTN 718 1e-51 83 421 2096 700046348H1 SATMON004 g683475 BLASTN 726 1e-51 87 422 2096 700968694H1 SATMONN04 g19060 BLASTN 306 1e-28 87 423 2096 700453783H1 SATMON029 g19060 BLASTN 197 1e-12 78 424 5587 LIB3062-053- LIB3062 g683475 BLASTN 1366 1e-111 91 Q1-K1-C4 425 5587 700087630H1 SATMON011 g683475 BLASTN 1021 1e-91 92 426 5587 700088983H1 SATMON011 g683475 BLASTN 708 1e-88 86 427 5587 700100889H1 SATMON009 g683475 BLASTN 1149 1e-87 89 428 5587 700470729H1 SATMON025 g683475 BLASTN 1006 1e-85 90 429 5587 700100883H1 SATMON009 g683475 BLASTN 839 1e-80 90 430 5587 700214072H1 SATMON016 g683475 BLASTN 910 1e-73 91 431 5587 700044664H1 SATMON004 g683475 BLASTN 587 1e-56 84 432 5587 LIB189-007-Q1- LIB189 g683475 BLASTN 403 1e-53 79 E1-A7 433 5587 700042060H1 SATMON004 g683475 BLASTN 734 1e-52 84

434 5587 LIB83-011-Q1- LIB83 g683475 BLASTN 310 1e-32 74 E1-A8 435 5587 700083223H1 SATMON011 g683475 BLASTN 310 1e-31 79 436 5587 700442503H1 SATMON026 g683475 BLASTN 458 1e-29 89 437 5587 700101540H1 SATMON009 g683475 BLASTN 310 1e-28 78 438 5587 700207961H1 SATMON016 g683475 BLASTN 304 1e-26 82 439 5587 700092960H1 SATMON008 g683475 BLASTN 310 1e-26 72 440 5587 700041761H1 SATMON004 g683475 BLASTN 301 1e-24 86 441 5587 700087935H1 SATMON011 g2244614 BLASTX 141 1e-12 100 442 5632 LIB3069-036- LIB3069 g510676 BLASTN 849 1e-76 81 Q1-K1-A5 443 5632 700243480H1 SATMON010 g16117 BLASTN 806 1e-73 89 444 5632 700198041H1 SATMON016 g16117 BLASTN 738 1e-52 90 445 5632 700097480H1 SATMON009 g19060 BLASTN 712 1e-50 80 446 5632 700088645H1 SATMON011 g19060 BLASTN 659 1e-46 79 447 5632 LIB3068-012- LIB3068 g19061 BLASTX 129 1e-35 90 Q1-K1-C6 448 5632 LIB3069-026- LIB3069 g19060 BLASTN 496 1e-34 75 Q1-K1-G11 449 5632 700081934H1 SATMON011 g16117 BLASTN 519 1e-34 89 450 5632 LIB3062-050- LIB3062 g16117 BLASTN 348 1e-29 83 Q1-K1-B3 451 5632 LIB3069-042- LIB3069 g19060 BLASTN 404 1e-25 72 Q1-K1-D6 452 5632 700095961H1 SATMON008 g19060 BLASTN 277 1e-23 68 453 5632 700091061H1 SATMON011 g19061 BLASTX 86 1e-10 64 454 5632 700224365H1 SATMON011 g19060 BLASTN 213 1e-10 63 455 5632 700089289H1 SATMON011 g19060 BLASTN 213 1e-10 63 456 5632 700094923H1 SATMON008 g19060 BLASTN 183 1e-9 63 457 5632 700094625H1 SATMON008 g19060 BLASTN 206 1e-9 63 458 5632 700093380H1 SATMON008 g19060 BLASTN 206 1e-9 63 459 5632 700093964H1 SATMON008 g19060 BLASTN 213 1e-9 69 460 5632 700083043H1 SATMON011 g19060 BLASTN 193 1e-8 63 461 5632 700095165H1 SATMON008 g510676 BLASTN 195 1e-8 65 462 5633 700094759H1 SATMON008 g510676 BLASTN 629 1e-43 86 463 5633 700094711H1 SATMON008 g16117 BLASTN 468 1e-32 89 464 9949 700213043H1 SATMON016 g16117 BLASTN 1192 1e-90 87 465 9949 700084670H1 SATMON011 g16117 BLASTN 1125 1e-85 88 466 9949 700213929H1 SATMON016 g16117 BLASTN 439 1e-27 84 MAIZE PUTATIVE PROTOCHLOROPHYLLIDE REDUCTASE 467 -700353742 700353742H1 SATMON024 g348717 BLASTN 512 1e-33 73 468 -700423111 700423111H1 SATMONN01 g348717 BLASTN 684 1e-48 72 469 15163 LIB3069-040- LIB3069 g348717 BLASTN 874 1e-64 70 Q1-K1-F9 470 15163 700623844H1 SATMON034 g348717 BLASTN 752 1e-53 69 471 15163 700623744H1 SATMON034 g348717 BLASTN 725 1e-51 70 472 15163 700623644H1 SATMON034 g348717 BLASTN 661 1e-46 72 473 15163 700612907H1 SATMON033 g348717 BLASTN 597 1e-40 73 474 15163 700612808H1 SATMON033 g348717 BLASTN 579 1e-39 74 475 15163 700623852H1 SATMON034 g348717 BLASTN 489 1e-30 67 476 15163 700475540H1 SATMON025 g348717 BLASTN 413 1e-26 65 477 22562 700571483H1 SATMON030 g348717 BLASTN 447 1e-26 72 478 30690 LIB3062-046- LIB3062 g348719 BLASTN 514 1e-31 69 Q1-K1-D4 479 30690 700425786H2 SATMONN01 g348720 BLASTX 168 1e-17 54 MAIZE COPROPORPHYRINOGEN OXIDASE 480 -L30623969 LIB3062-019- LIB3062 g1213067 BLASTX 123 1e-25 76 Q1-K1-A9 481 26808 LIB3062-022- LIB3062 g1213067 BLASTX 195 1e-35 89 Q1-K1-A9 482 26808 LIB36-007-Q1- LIB36 g414665 BLASTN 235 1e-8 85 E1-H7 483 5948 700614009H1 SATMON033 g1212993 BLASTN 1318 1e-101 87 484 5948 LIB3078-027- LIB3078 g1212993 BLASTN 1185 1e-89 83 Q1-K1-C2 485 5948 700207069H1 SATMON003 g1212993 BLASTN 1003 1e-82 81 486 5948 701183985H1 SATMONN06 g1212993 BLASTN 1064 1e-79 88 487 5948 700043235H1 SATMON004 g1212993 BLASTN 944 1e-69 85 488 5948 700237643H1 SATMON010 g1212993 BLASTN 920 1e-67 85 489 5948 700167142H1 SATMON013 g1212993 BLASTN 832 1e-60 85 490 98 LIB3062-011- LIB3062 g1212993 BLASTN 1515 1e-120 85 Q1-K1-B9 491 98 700089965H1 SATMON011 g1212993 BLASTN 1129 1e-85 85 492 98 700473370H1 SATMON025 g1212993 BLASTN 812 1e-79 83 493 98 700018492H1 SATMON001 g1212993 BLASTN 650 1e-45 87 494 98 700336060H1 SATMON019 g1212993 BLASTN 423 1e-26 83 MAIZE PROTOPORPHYRINOGEN OXIDASE 495 13987 700397414H1 SATMONN01 g1877018 BLASTX 152 1e-13 72 496 13987 700377840H1 SATMON019 g2370333 BLASTX 115 1e-8 75 497 21128 700087081H1 SATMON011 g1183006 BLASTN 851 1e-62 75 498 21128 700222959H1 SATMON011 g1183006 BLASTN 551 1e-47 74 499 8675 LIB3062-009- LIB3062 g3093409 BLASTN 1093 1e-82 72 Q1-K1-F6 MAIZE UROPORPHYRINOGEN DECARBOXYLASE 500 -700210906 700210906H1 SATMON016 g1009429 BLASTX 172 1e-25 50 501 -700334993 700334993H1 SATMON019 g1009427 BLASTN 515 1e-70 84 502 -700432067 700432067H1 SATMONN01 g216564 BLASTX 123 1e-14 39 503 -L1891364 LIB189-002-Q1- LIB189 g1009427 BLASTN 914 1e-78 85 E1-E8 504 -L30625966 LIB3062-056- LIB3062 g1322019 BLASTX 660 1e-102 100 Q1-K1-D10 505 -L30626254 LIB3062-058- LIB3062 g1009427 BLASTN 516 1e-32 81 Q1-K1-D9 506 -L30783694 LIB3078-054- LIB3078 g1009427 BLASTN 1355 1e-104 84 Q1-K1-D9 507 30392 700090031H1 SATMON011 g1009427 BLASTN 794 1e-79 92 508 30392 LIB3062-053- LIB3062 g1009427 BLASTN 762 1e-54 89 Q1-K1-D9 509 30392 LIB3069-027- LIB3069 g1009427 BLASTN 664 1e-44 83 Q1-K1-G9 MAIZE PUTATIVE UROPORPHYRINOGEN DECARBOXYLASE 510 -700799143 700799143H1 SATMON036 g48040 BLASTX 128 1e-21 47 MAIZE PORPHOBILINOGEN SYNTHASE 511 -700082696 700082696H1 SATMON011 g1041422 BLASTN 544 1e-64 86 512 -700421637 700421637H1 SATMONN01 g1041422 BLASTN 591 1e-57 85 513 10331 700049523H1 SATMON003 g1041422 BLASTN 694 1e-73 84 514 10331 700214149H1 SATMON016 g1041422 BLASTN 986 1e-73 84 515 6252 LIB3060-049- LIB3060 g1041422 BLASTN 1296 1e-119 85 Q1-K1-D11 516 6252 700104193H1 SATMON010 g1041422 BLASTN 1117 1e-84 87 517 6252 700043614H1 SATMON004 g1041422 BLASTN 1081 1e-81 87 518 6252 700104333H1 SATMON010 g1041422 BLASTN 757 1e-76 86 519 6252 700099573H1 SATMON009 g1041422 BLASTN 969 1e-71 85 520 6252 LIB189-034-Q1- LIB189 g1041422 BLASTN 829 1e-64 82 E1-G11 521 6252 700150031H1 SATMON007 g1041422 BLASTN 715 1e-50 80 522 6252 700150305H1 SATMON007 g1041422 BLASTN 494 1e-32 88 523 6664 700098341H1 SATMON009 g1041422 BLASTN 861 1e-62 80 524 6664 700097010H1 SATMON009 g1041422 BLASTN 861 1e-62 80 525 6664 700150830H1 SATMON007 g1041422 BLASTN 655 1e-45 78 526 6664 700088427H1 SATMON011 g1041422 BLASTN 598 1e-41 85 527 6664 700216648H1 SATMON016 g1041422 BLASTN 586 1e-40 78 528 6664 700150750H1 SATMON007 g1041422 BLASTN 562 1e-38 77 529 6664 700089504H1 SATMON011 g1041422 BLASTN 349 1e-35 81 530 6664 700150781H1 SATMON007 g1041422 BLASTN 473 1e-30 71 531 6664 700071849H1 SATMON007 g1041423 BLASTX 158 1e-14 66 MAIZE HYDROXYMETHYLBILANE SYNTHASE 532 -700042853 700042853H1 SATMON004 g2661765 BLASTN 1319 1e-101 96 533 -700209530 700209530H1 SATMON016 g2661765 BLASTN 1046 1e-96 91 534 -L30784536 LIB3078-039- LIB3078 g2661765 BLASTN 980 1e-73 81 Q1-K1-D10 535 18 700434552H1 SATMONN01 g2661765 BLASTN 819 1e-59 77 536 18 700621233H1 SATMON034 g2661765 BLASTN 606 1e-44 85 537 18 700621333H1 SATMON034 g2661765 BLASTN 607 1e-41 88 538 22370 LIB3078-049- LIB3078 g2661765 BLASTN 1197 1e-91 93 Q1-K1-D11 539 22370 LIB3078-007- LIB3078 g313723 BLASTN 745 1e-72 71 Q1-K1-F2 540 22370 700223478H1 SATMON011 g313723 BLASTN 712 1e-50 72 541 22370 700216196H1 SATMON016 g313723 BLASTN 508 1e-45 74 542 22370 700551081H1 SATMON022 g2661765 BLASTN 328 1e-36 91 MAIZE GLUTAMATE-1-SEMIALDEHYDE 2,1-AMINOMUTASE 543 -L841669 LIB84-026-Q1- LIB84 g506383 BLASTX 164 1e-42 69 E1-D6 544 11095 LIB3078-054- LIB3078 g556018 BLASTN 1229 1e-93 81 Q1-K1-E2 545 11095 LIB83-005-Q1- LIB83 g556018 BLASTN 1169 1e-88 80 E1-B11 546 11095 700101450H1 SATMON009 g556018 BLASTN 1024 1e-76 81 547 11095 700342512H1 SATMON021 g556018 BLASTN 995 1e-74 82 548 11095 700265085H1 SATMON017 g556018 BLASTN 996 1e-74 82 549 11095 700154602H1 SATMON007 g556018 BLASTN 491 1e-58 82 550 11095 700154908H1 SATMON007 g556018 BLASTN 722 1e-58 81 551 11095 700017624H1 SATMON001 g556018 BLASTN 775 1e-55 83 552 11095 700018108H1 SATMON001 g556018 BLASTN 758 1e-54 84 553 11095 700443671H1 SATMON027 g556018 BLASTN 662 1e-46 73 554 11095 700442812H1 SATMON026 g556018 BLASTN 578 1e-39 80 555 11095 700343762H1 SATMON021 g556018 BLASTN 565 1e-38 80 556 11095 700094251H1 SATMON008 g19873 BLASTX 167 1e-16 89 557 11225 LIB3060-054- LIB3060 g556018 BLASTN 817 1e-69 77 Q1-K1-C12 558 11225 700100123H1 SATMON009 g556018 BLASTN 787 1e-56 85 559 11225 700405062H1 SATMON027 g556018 BLASTN 453 1e-34 75 560 11225 700219159H1 SATMON011 g556018 BLASTN 328 1e-26 74 561 11225 700209352H1 SATMON016 g506383 BLASTX 174 1e-17 70 562 11225 700053276H1 SATMON008 g506383 BLASTX 131 1e-10 96 563 11225 700156122H2 SATMON007 g506383 BLASTX 120 1e-9 100 564 15553 700084357H1 SATMON011 g556018 BLASTN 1104 1e-83 80 565 15553 700441108H1 SATMON026 g556018 BLASTN 1071 1e-80 86 566 15553 700441006H1 SATMON026 g556018 BLASTN 1062 1e-79 86 567 15553 700087059H1 SATMON011 g556018 BLASTN 421 1e-26 82 568 20096 700089246H1 SATMON011 g556018 BLASTN 474 1e-49 82 569 20096 700171369H1 SATMON013 g556018 BLASTN 560 1e-37 78 MAIZE GLUTAMATE tRNA LIGASE 570 -700614160 700614160H1 SATMON033 g31958 BLASTX 113 1e-10 56 571 -L1892744 LIB189-012-Q1- LIB189 g31958 BLASTX 146 1e-28 50 E1-F8 572 -L1894036 LIB189-029-Q1- LIB189 g157564 BLASTX 143 1e-28 62 E1-B1 573 12385 LIB3067-058- LIB3067 g2995455 BLASTX 416 1e-70 69 Q1-K1-H9 574 13776 700344387H1 SATMON021 g157564 BLASTX 108 1e-19 57 575 21786 700221143H1 SATMON011 g157564 BLASTX 287 1e-32 59 576 26250 LIB3069-031- LIB3069 g2995455 BLASTX 166 1e-43 74 Q1-K1-E6 577 3350 LIB3069-025- LIB3069 g157564 BLASTX 232 1e-44 46 Q1-K1-F6 578 3350 700072785H1 SATMON007 g157564 BLASTX 249 1e-26 45 579 3350 700049536H1 SATMON003 g157564 BLASTX 227 1e-24 50 580 3350 700077013H1 SATMON007 g157564 BLASTX 232 1e-24 49 581 3350 700209830H1 SATMON016 g157564 BLASTX 210 1e-22 52 582 3350 700168681H1 SATMON013 g157564 BLASTX 156 1e-14 40 583 5345 LIB3059-036- LIB3059 g2995455 BLASTX 148 1e-28 67 Q1-K1-G10 584 9230 LIB143-053-Q1- LIB143 g31958 BLASTX 341 1e-55 58 E1-G8 585 9230 700331892H1 SATMON019 g157564 BLASTX 162 1e-31 55 MAIZE GLUTAMYL-tRNA REDUCTASE 586 -700094403 700094403H1 SATMON008 g1039331 BLASTN 740 1e-63 83 587 -700151003 700151003H1 SATMON007 g1039331 BLASTN 885 1e-64 87 588 -700167046 700167046H1 SATMON013 g1039331 BLASTN 772 1e-64 89 589 -L30661635 LIB3066-003- LIB3066 g1666078 BLASTN 298 1e-18 77 Q1-K1-A8 590 -L30661878 LIB3066-012- LIB3066 g2967440 BLASTN 485 1e-29 85 Q1-K1-F3 591 -L362024 LIB36-016-Q2- LIB36 g2920319 BLASTN 170 1e-9 70 E2-H11 592 22014 700045741H1 SATMON004 g1039331 BLASTN 921 1e-67 82 593 22014 700214783H1 SATMON016 g1039331 BLASTN 860 1e-62 83 594 22618 700086081H1 SATMON011 g1039331 BLASTN 1064 1e-79 82 595 22618 700104481H1 SATMON010 g1039331 BLASTN 955 1e-70 81 596 22618 700356789H1 SATMON024 g1039331 BLASTN 644 1e-44 86 597 30084 LIB3062-026- LIB3062 g2920319 BLASTN 1043 1e-78 87 Q1-K1-H5 598 30084 701179026H1 SATMONN05 g1039331 BLASTN 753 1e-72 88 599 6787 LIB36-021-Q1- LIB36 g1039331 BLASTN 1281 1e-97 87 E1-D9 600 6787 701163632H1 SATMONN04 g1039331 BLASTN 941 1e-79 86 601 6787 700162337H1 SATMON012 g1039331 BLASTN 901 1e-66 84 602 6787 700100879H1 SATMON009 g1039331 BLASTN 608 1e-65 86 603 6787 700425112H1 SATMONN01 g1039331 BLASTN 230 1e-9 81 604 9690 LIB3078-023- LIB3078 g1039331 BLASTN 1755 1e-137 88 Q1-K1-F12 605 9690 700097404H1 SATMON009 g1039331 BLASTN 1311 1e-100 89 606 9690 700099954H1 SATMON009 g1039331 BLASTN 1132 1e-92 90 607 9690 700213724H1 SATMON016 g1039331 BLASTN 1211 1e-92 89 608 9690 700468009H1 SATMON025 g1039331 BLASTN 1187 1e-90 88 609 9690 700042554H1 SATMON004 g1039331 BLASTN 931 1e-68 88 MAIZE Mg-CHELATASE 610 -700042626 700042626H1 SATMON004 g861198 BLASTN 489 1e-31 67 611 -700045010 700045010H1 SATMON004 g861198 BLASTN 824 1e-59 80 612 -700046489 700046489H1 SATMON004 g861198 BLASTN 246 1e-11 93 613 -700090155 700090155H1 SATMON011 g2239151 BLASTX 226 1e-24 84 614 -700100867 700100867H1 SATMON009 g2239150 BLASTN 944 1e-69 81 615 -700152555 700152555H1 SATMON007 g861198 BLASTN 800 1e-57 83 616 -700166615 700166615H1 SATMON013 g861198 BLASTN 438 1e-27 88 617 -700214027 700214027H1 SATMON016 g847872 BLASTN 683 1e-67 80 618 -700216016 700216016H1 SATMON016 g861198 BLASTN 408 1e-24 78 619 -700219682 700219682H1 SATMON011 g2318116 BLASTN 616 1e-42 70 620 -L30606220 LIB3060-019- LIB3060 g861199 BLASTX 106 1e-32 67 Q1-K1-D2 621 15513 700379674H1 SATMON021 g2239151 BLASTX 138 1e-11 87 622 15984 700223402H1 SATMON011 g2318116 BLASTN 830 1e-60 78 623 15984 701185568H1 SATMONN06 g2318116 BLASTN 787 1e-56 79 624 15984 701185572H1 SATMONN06 g2239150 BLASTN 528 1e-48 78 625 15984 700257978H1 SATMON017 g2239150 BLASTN 454 1e-33 72 626 19005 700025578H1 SATMON004 g861198 BLASTN 874 1e-63 84 627 19005 700442062H1 SATMON026 g861198 BLASTN 226 1e-23 81 628 19969 700100921H1 SATMON009 g847872 BLASTN 1183 1e-89 87 629 19969 700422986H1 SATMONN01 g847872 BLASTN 598 1e-75 88 630 19969 700216568H1 SATMON016 g847872 BLASTN 822 1e-59 89

631 21239 LIB36-010-Q1- LIB36 g847872 BLASTN 1338 1e-139 88 E1-H9 632 21239 700053384H1 SATMON009 g847872 BLASTN 596 1e-85 88 633 21239 700043650H1 SATMON004 g847872 BLASTN 1122 1e-84 90 634 21239 700160759H1 SATMON012 g847872 BLASTN 760 1e-74 89 635 29840 LIB84-024-Q1- LIB84 g861198 BLASTN 1195 1e-90 80 E1-B8 636 29840 700046445H1 SATMON004 g861198 BLASTN 898 1e-65 82 637 29840 700099676H1 SATMON009 g861198 BLASTN 755 1e-54 79 638 29840 700216064H1 SATMON016 g861198 BLASTN 433 1e-25 74 639 3221 700342738H1 SATMON021 g2239150 BLASTN 678 1e-74 82 640 3221 700090050H1 SATMON011 g2239150 BLASTN 556 1e-66 80 641 3221 700259555H1 SATMON017 g2239151 BLASTX 215 1e-27 72 642 5373 LIB3078-054- LIB3078 g861198 BLASTN 1220 1e-92 82 Q1-K1-A2 643 5373 700097131H1 SATMON009 g861198 BLASTN 1071 1e-80 82 644 5373 700043508H1 SATMON004 g861198 BLASTN 1017 1e-75 83 645 5373 700046404H1 SATMON004 g861198 BLASTN 988 1e-73 83 646 5373 700431104H1 SATMONN01 g861198 BLASTN 769 1e-65 82 647 5373 700043839H1 SATMON004 g861198 BLASTN 747 1e-53 83 648 5373 700433984H2 SATMONN01 g861198 BLASTN 493 1e-32 76 649 5953 LIB3078-052- LIB3078 g861198 BLASTN 1148 1e-92 81 Q1-K1-A9 650 5953 700045450H1 SATMON004 g861198 BLASTN 1005 1e-74 83 651 5953 700041735H1 SATMON004 g861198 BLASTN 969 1e-71 83 652 5953 LIB83-001-Q1- LIB83 g861198 BLASTN 706 1e-58 83 E1-F5 MAIZE FERROCHELATASE 653 -700214704 700214704H1 SATMON016 g2429617 BLASTN 822 1e-59 85 654 -700239147 700239147H1 SATMON010 g439480 BLASTN 905 1e-66 87 655 -700382669 700382669H1 SATMON024 g439480 BLASTN 1079 1e-81 87 656 -700441938 700441938H1 SATMON026 g2429617 BLASTN 766 1e-55 84 657 -700579576 700579576H1 SATMON031 g439480 BLASTN 739 1e-52 75 658 -L1892866 LIB189-014-Q1- LIB189 g2429617 BLASTN 248 1e-15 78 E1-E6 659 -L832454 LIB83-005-Q1- LIB83 g2460250 BLASTN 303 1e-35 88 E1-F4 660 11690 700151225H1 SATMON007 g2429617 BLASTN 1083 1e-81 91 661 11690 700106040H1 SATMON010 g2429617 BLASTN 627 1e-59 90 662 11690 700167395H1 SATMON013 g2429617 BLASTN 637 1e-44 91 663 14766 LIB143-007-Q1- LIB143 g439480 BLASTN 580 1e-56 77 E1-D5 664 14766 700263637H1 SATMON017 g439480 BLASTN 259 1e-10 72 665 14766 LIB36-007-Q1- LIB36 g2460250 BLASTN 243 1e-8 72 E1-C9 666 16136 700354263H1 SATMON024 g439480 BLASTN 987 1e-76 87 667 16136 700222207H1 SATMON011 g439480 BLASTN 903 1e-66 82 668 17054 700158001H1 SATMON012 g439480 BLASTN 1091 1e-82 90 669 17054 700157069H1 SATMON012 g439480 BLASTN 1096 1e-82 90 670 17054 700453159H1 SATMON028 g439480 BLASTN 816 1e-73 88 671 394 700622934H1 SATMON034 g439480 BLASTN 763 1e-84 86 672 394 700621391H1 SATMON034 g439480 BLASTN 775 1e-82 86 673 394 700098357H1 SATMON009 g2460251 BLASTX 155 1e-13 83 674 9731 LIB3078-007- LIB3078 g2429617 BLASTN 1299 1e-123 85 Q1-K1-B10 675 9731 700355823H1 SATMON024 g2429617 BLASTN 826 1e-84 87 676 9731 700208201H1 SATMON016 g2429617 BLASTN 702 1e-64 82 677 9731 700167772H1 SATMON013 g2429617 BLASTN 648 1e-45 88 *Table Headings Cluster ID A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a "singleton"), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. The cluster ID entries in the table refer to the cluster with which the particular clone in each row is associated. Clone ID The clone ID number refers to the particular clone in the PhytoSeq database. Each clone ID entry in the table refers to the clone whose sequence is used for (1) the sequence comparison whose scores are presented and/or (2) assignment to the particular cluster which is presented. Note that a clone may be included in this table even if its sequence comparison scores fail to meet the minimum standards for similarity. In such a case, the clone is included due solely to its association with a particular cluster for which sequences of one or more other member clones possess the required level of similarity. Library The library ID refers to the particular cDNA library from which a given clone is obtained. Each cDNA library is associated with the particular tissue(s), line(s) and developmental stage(s) from which it is isolated. NCBI gi Each sequence in the GenBank public database is arbitrarily assigned a unique NCBI gi (National Center for Biotechnology Information GenBank Identifier) number. In this table, the NCBI gi number which is associated (in the same row) with a given clone refers to the particular GenBank sequence which is used in the sequence comparison. This entry is omitted when a clone is included solely due to its association with a particular cluster. Method The entry in the "Method" column of the table refers to the type of BLAST search that is used for the sequence comparison. "CLUSTER" is entered when the sequence comparison scores for a given clone fail to meet the minimum values required for significant similarity. In such cases, the clone is listed in the table solely as a result of its association with a given cluster for which sequences of one or more other member clones possess the required level of similarity. Score Each entry in the "Score" column of the table refers to the BLAST score that is generated by sequence comparison of the designated clone with the designated GenBank sequence using the designated BLAST method. This entry is omitted when a clone is included solely due to its association with a particular cluster. If the program used to determine the hit is HMMSW then the score refers to HMMSW score. P-Value The entries in the P-Value column refer to the probability that such matches occur by chance. % Ident The entries in the "% Ident" column of the table refer to the percentage of identically matched nucleotides (or residues) that exist along the length of that portion of the sequences which is aligned by the BLAST comparison to generate the statistical scores presented. This entry is omitted when a clone is included solely due to its association with a particular cluster.

Sequence CWU 1

6771257DNAGlycine max 1tgctgcttct ggaaattttc attggaattt tgaagatgtt gctaaatcaa ttgtgtgcat 60gatgatgtct ggcccattct tgacaggata tacccagact atgaatgatt ggtacgaccg 120agaaattgat gcaataaatg aaccttatag accaattcct tctggggcaa tatctgagaa 180tgaggtaatc actcaaatat gggtgttgct gcttggtggt ctttctctgg ctggtatatt 240ggacatatgg gcagggc 2572272DNAGlycine maxunsure at all n locations 2cacatgtaag catctcaagc tctgctgaat cttcaatggc ttctctactc aacatggttt 60ctgttccatc aagaatatca ccaagctcac acacgagaac cacttcaang caatctcgaa 120ctgttttgcc accattttct gtctcatttt ccaggaggag attatcaatt agagcaacag 180aaactgatac taatgaagtt caatctcagg cgccgggtac agcaccatca aaagatggtt 240caagcttcaa ccagctcctt ggtattaaag ga 2723156DNAGlycine max 3aagaaacaaa taagtggaag attcgtcttc aacttacaaa gccagtcact tggcctccat 60taatttgggg tgtagtttgt ggagctgctg cttctggaaa ttttcattgg aattttgaga 120tgttgctaaa tcaattgtgt gcatgatgat gtctgg 1564348DNAGlycine max 4agtacggctg cgagaagacg acagaagggg aaggcatctt caagctctga atctgcaatg 60gcttctctac tcaacatggt ttcggttcca ccaagaatat caccaacctc acacaccaga 120atcgcttcgc ttcaagctcg acccgttttg ccaccctttt ctgtctcatt ttccaggagg 180agactatcaa ttagagcaac agaaactgat accaatgaag ttcaatctca ggcaccgggt 240gcagcgccat ctaaagatgg ttcaagcttc aatcagcttc ttggtatcaa aggagctgcc 300caagaaacaa ataaatggaa aattcgtctt caactcacaa agcctgtc 3485245DNAGlycine maxunsure at all n locations 5ctctgaatct gcaatggctt ctctactcaa catggtttcg gttncaccaa gactatcact 60cnnctcacac accagaatcg cttcgcttca agctcgaccc gtttgccacc cttttctgtc 120tcattttcca ggaggagact atcaattaga gcaacagaaa ctgataccaa tgaagttcaa 180tctcaggcac cgggtgcagc gccatctaaa gatggttcaa gcttcaatca gcttcttggt 240atcaa 2456268DNAGlycine max 6tggcatcttc aagctctgaa tctgcaatgg cttctctact caacatggtt tcggttccac 60caagaatatc accaacctca cacaccagaa tcgcttcgct tcaagctcga cccgttttgc 120cacccttttc tgtctcattt tccaggagga gactatcaat tagagcaaca gaaactgata 180ccaatgaagt tcaatctcag gcaccgggtg cagcgccatc taaagatggt tcaagcttca 240atcagcttct tggtatcaaa ggagctgc 2687278DNAGlycine max 7cggctgcgag aagacgacag aagggctcag agtactgtta ttgaaaggca aaggacaata 60gagtatacct gaagccctag agccctatcc ccttcaacac ttttgaagtc attgacaata 120gcaattccca actgcaatgt gatttaacaa caacattaat aaccattttt atttgacata 180ttatcatatt catatccaac aaaatgtcat gaagaatata ttacatactc cagctatgct 240gtataggagt gtgagaacaa ttatatctgg tgtaagag 2788248DNAGlycine max 8cggctgcgag aagacgacag aagggctcag agtactgtta ttgaaaggca aaggacaata 60gagtatacct gaagccctag agccctatcc ccttcaacac ttttgaagtc attgacaata 120gcaattccca actgcaatgt gatttaacaa caacattaat aaccattttt atttgacata 180ttatcatatt catatccaac aaaatgtcat gaagaatata ttacatactc cagctatgct 240gtatagga 2489258DNAGlycine maxunsure at all n locations 9gncanctgct angganccta cntncactgg cngagggctt tgaacttagc ctnnnggaca 60aatcatctng ggcatttcct cctctcgcgc cngttgctng aggacttgga naaatncgag 120tacccttcaa aggcttgatn atcgtaggnt cacacgacag ggnacacaaa cacattggct 180ggtaatgtac ctcccaaggc gaaccttggn ggacttgagg ggacttcagg gtggtttgaa 240tgggctaaag agctcagc 25810270DNAGlycine max 10gtcaatttgt tgataacttt aggcaatcag gccggccact ggatgtgctt gtttgcaatg 60ctgcggttta cttgccaact gccagggaac ctacatatac tgctgatggc tttgaactca 120gtgttggaac caaccatctc gggcatttcc tcctttcgcg ccttttgctt gacgacttga 180acaaatctga ctacccttcg aagcggttga tcatgtaggc tcaatcacag gaaacaccaa 240cacattggct ggaatgtgcc acccaggcta 27011258DNAGlycine maxunsure at all n locations 11caggaaacac caacacattg gctggaaatg tgccacccaa ggctaacctt ggtgacatga 60ggggactagc tggaggcttg aatgggctaa acacttcagc catgatagat ggaggatcct 120ttgacggcgc taaggcatac aaggacagca aagtctgcaa catgcttaca atgccagaat 180tccaacagga ggtcccngtt ganaccnngg natnacatnt gcncccntan cccngggttn 240ttcncccaaa ngggnttt 25812270DNAGlycine max 12gacggcgcta aggcatacaa ggacagcaaa gtctgcaaca tgcttacaat gcaagaattc 60cacagaagat accatgatga aactgggatc acatttgctt ccctttaccc aggttgcatc 120gccacaacag gcttgttcag agagcacatt cccttgttca gacttctctt ccctccattc 180caaaagtaca taaccaaggg ctttgtctca gaagatgaat caggaaagag acttgcacag 240gttgtgagtg atccaagcct aacaaaatca 27013262DNAGlycine max 13caggctgctt ctttccccat tgctaaagag ggaaagtctg gtgtttctct caggtacacc 60acaatgttcg gtgtttcatt gtcggatact ctcaaatctg acgctcagct tttcctcatt 120gacatgcaaa gaaacaccaa caccttggct ggacatgtgc cacccaaggc taaccttggt 180gacttgaggg gactagctgg aggcttgaat gggctaaaca cttcagccat gatagatgga 240ggatcctttg atggcaccaa gg 26214279DNAGlycine maxunsure at all n locations 14ccatttgctt ccctttaccc cggttgcatt gccacaacag gcctgttcag agagcacatt 60cccttgttca naactctgtt ccctccattc cagaagtaca taaccaaagg ctatgtctca 120gaagatgaag caggaaagag acttgctcag gttgtaagtg atccaagcct aacaaaatct 180ggtgtttact ggagctggaa caaagcatca gcttcgtttg aaaaccagtt gtctcaggag 240gccagtgata cagagaaggc tcgtaagatc tgggagnta 27915346DNAGlycine max 15aaacaaagga cccagtttta catttttttt tgttcctgag ttccaatggc tcttcaggct 60gcttccttgg tttctgcttc tttttctatt gctaaagagg gaaagtctgg tgtatctctc 120agggacacca caatgtttgg tgtttcattg tcggatactc tcaaatctga cttcagctct 180ccctcatcga cttgcaaaag ggaattccaa caaaaatttg gccctttgag ggttcagtca 240gtggcaacaa caactccagg agtcaccaag gcttcaccag aaggcaagaa aactttgagg 300aaaggcagtg ttattatcac tggggcttcc tctggattag gctggc 34616256DNAGlycine max 16ctaaaacaaa ggacccagtt ttacattttt ttcctgagtt ccaatggctc ttcaggctgc 60ttccttggtt tctgcttctt tttctattgc taaagaggga aagtctggtg tatctctcag 120ggacaccaca atgtttggtg tttcattgtc ggatactctc aaatctgact tcagctctcc 180ctcatcgact tgcaaaaggg aattccaaca aaaatttggc cctttgaggg ttcagtcagt 240ggcaacaaca actcca 25617269DNAGlycine max 17cagttttaca tttttttttg ttcctgagtt ccaatggctc ttcaggctgc ttccttggtt 60tctgcttctt tttctattgc taaagaggga aagtctggtg tatctctcag ggacaccaca 120atgtttggtg tttcattgtc ggatactctc aaatctgact tcagctctcc ctcatcgact 180tgcaaaaggg aattccaaca aaaatttggc cctttgaggg ttcagtcagt ggcaacaaca 240actccaggag tcaccaaggc ttcaccaga 26918358DNAGlycine maxunsure at all n locations 18gaaacattct aaaacaaagg acccagtttt acatttttnt ttgttcctga gttccaatgg 60ctcttcaggc tgcttccttg gtttctgctt ctttttctat tgctaaagag ggaaagtctg 120gtgtatctct cagggacacc acaatgtttg gtgtttcatt gtcggatact ctcaaatctg 180acttcagctc tccctcatcg acttgcaaaa gggaattcca acaaaaattt ggccctttga 240gggttcagtc agtggcaaca acaactccag gagtcaccaa ggttcaccag aaggcaagaa 300ctttgaggaa ggcagtgnta taccatgggg cttcctctgg attagcctgg cactgcta 35819264DNAGlycine max 19aaacattcta aaacaaagga cccagtttta catttttttt tgttcctgag ttccaatggc 60tcttcaggct gcttccttgg tttctgcttc tttttctatt gctaaagagg gaaagtctgg 120tgtatctctc agggacacca caatgtttgg tgtttcattg tcggatactc tcaaatctga 180cttcagctct ccctcatcga cttgcaaaag ggaattccaa caaaaatttg gccctttgag 240ggttcagtca gtggcaacaa caac 26420253DNAGlycine max 20acattctaaa acaaaggacc cagttttaca tttgtttttg ttcctgagtt ccaatggctc 60ttcaggctgc ttccttggtt tctgcttctt tttctattgc taaagaggga aagtctggtg 120tatctctcag ggacaccaca atgtttggtg tttcattgtc ggatactctc aaatctgact 180tcagctctcc ctcatcgact tgcaaaaggg aattccaaca aaaatttggc cctttgaggg 240ttcagtcagt ggc 25321256DNAGlycine max 21acattctaaa acaaaggacc cagttttaca tttttgtttg ttcctgagtt ccaatggctc 60ttcaggctgc ttccttggtt tctgcttctt tttctattgc taaagaggga aagtctggtg 120tatctctcag ggacaccaca atgtttggtg tttcattgtc ggatactctc aaatctgact 180tcagctctcc ctcatcgact tgcaaaaggg aattccaaca aaaatttggc cctttgaggg 240ttcagtcagt ggcaac 25622277DNAGlycine max 22atttttattt gttcctgagt tccaatggct cttcaggctg cttccttggt ttctgcttct 60ctttctattg ctaaagaggg aaagtctggt gtatctctca gggactccac aatgtttggt 120gtttcattgt cggatactct caaatctgac ttcagctctc tctcatcgac ttgcaaaagg 180gaattccaac aaaaatttgg cccgttaagg gttcagtcag tggcaacaac aactccagga 240gtcaccaagg cttcaccaga aggcgatgaa atttgag 27723256DNAGlycine max 23gaaacattct aaaacaaagg acccagtttt acattttttt tgttcctgag ttccaatggc 60tcttcaggct gcttccttgg tttctgcttc tttttctatt gctaaagagg gaaagtctgg 120tgtatctctc agggacacca caatgtttgg tgtttcattg tcggatactc tcaaatctga 180cttcagctct ccctcatcga cttgcaaaag ggaattccaa caaaaatttg gccctttgag 240ggttcagtca gtggca 25624269DNAGlycine max 24gttttacatt ttttttttgt tcctgagttc caatggctct tcaggctgct tccttggttt 60ctgcttcttt ttctattgct aaagagggaa agtctggtgt atctctcagg gacaccacaa 120tgtttggtgt ttcattgtcg gatactctca aatctgactt cagctctccc tcatcgactt 180gcaaaaggga attccaacaa aaatttggcc ctttgagggt tcagtcagtg gcaacaacaa 240ctccaggagt caccaaggct tcaccagaa 26925251DNAGlycine max 25gcttctttcc ccattgctaa agagggaaag tctggtgttt ctctcaggta caccacaatg 60ttcggtgttt cattgtcgga tactctcaaa tcagacttca gcttttcctc attgacatgc 120aaaagggaat tccaacaaaa aattggccct ttgagggttc agtcagtggc aacaaccact 180ccaggagtca ccaaggcttc accagaaggc aagaaaactt tgaggaaagg cagtgttatt 240gtcactgggc t 25126246DNAGlycine max 26ggctcgagaa cattctaaaa caaaggaccc aattttacat ttttttcact tcctgagttc 60caatggctct tcaggctgct tccttggttt ctgcttcttt ttctattgct aaagagggaa 120agtctggtgt atctctcagg gacaccacaa tgtttggtgt ttcattgtcg gatactctca 180aatctgactt cagctctccc tcatcgactt gcaaaaggga attccaacaa aaatttggcc 240ctttga 24627254DNAGlycine max 27gaaacattct aaaacaaagg acccagtttt acattttttt ttgttcctga gttccaatgg 60ctcttcaggc tgcttccttg gtttctgctt ctttttctat tgctaaagag ggaaagtctg 120gtgtatctct cagggacacc acaatgtttg gtgtttcatt gtcggatact ctcaaatctg 180acttcatctc tccctcatcg acttgcaaaa gggaattcca acaaaaattt ggccctttga 240gggttcagtc agtg 25428259DNAGlycine maxunsure at all n locations 28aaacaaagga cccagtttta catttttttt tgttcctgag ttccaatggc tcttcaggct 60gcttccttgg tttctgcttc tttttctatt gctaaagagg gaaagtctgg tgtatctctc 120agggacacca caatgtttgg tgtttcattg tcggatactc tcaaatctna cttcagctct 180ccctcatcga cttgcaaaag ggaattccaa canaaatttg gccccgggtt cagtcagtgg 240naacaacaac ncgnggagt 25929249DNAGlycine maxunsure at all n locations 29aaacattcta aaacaaagga cccagtttta catttttntt tgttcctgag ttccaatggc 60tnctccaggc tgcttccttg gtttctgctt cttttnctat tgttaaagag ggaaagttct 120ggtgtatctc tcagggacac cacnatgttt ggtgtttcat tgtcggatac tctcaaatct 180gacttcagct ctccctcatc gacttgcaaa agggaattcc aacanaaatt tggccctttg 240agggttcag 24930230DNAGlycine max 30gaaacattct aaaacaaagg acccagtttt acattttttt ttgttcctga gttccaatgg 60ctcttcaggc tgcttcctgt ggtttctgct tctttttcta ttgctaaaga gggaaagtct 120ggtgtatctc tcagggacac cacaatgttt ggtgtttcat tgtcggatac tctcaaatct 180gacttcagct ctccctcatc gacttgcaaa agggaattcc aacaaaaatt 23031445DNAGlycine maxunsure at all n locations 31gcgagaagac gacagaaggg gtctcagaag atgaagcagg aaagagactt gctcaggttg 60taagtgatcc aagcctaaca aaatctggtg tttactggag ctgaaacaaa gcatcagctt 120cgtttgaaaa ccagttgtct caggaggcca gtgatacaga gaaggctcgt aagatctggg 180agattagtga gaaacttgtt ggttttgcct aagtgggagg agcctccaac atcccatgtt 240gttctagaga ccttgcactt gcatggagga agaaaatgat gtctcaaaag agtggataga 300taacatccta tcattttgaa tgcattgatg ttgttttgtt agctaggagc ttctttgctt 360tgatgtaagg tgtcaatggc tttttgtgaa tcaagactca ataaaatcat tcagccatgt 420gggtgtggtg aagttgctca taana 44532256DNAGlycine max 32attgctcagg ttgtaagtga tccaagccta acaaaatctg gtgtttactg gagctggaac 60aaagcatcag cttcgtttga aaaccagttg tctcaggagg ccagtgatac agagaaggct 120cgtaagatct gggagattag tgagaaactt gttggttttg cctaagtggg aggagcctcc 180aacatcccat gttgttctag agaccttgca cttgcatgga ggaagaaaat gacgtctcaa 240aagagtggat agataa 25633259DNAGlycine maxunsure at all n locations 33ggctaaacag ctcagccatg attgatggtg gagacttcga tggtgccaag gcgtacaagg 60acagcaaagt ctgcaatatg ctcacaatgc aagaattcca cagacgattc catgaggaaa 120ctggaatcac atttgcttcc ctttaccccg gttgcattgc cacaacaggc ctgttcagag 180agcacttccc ttgttcagaa actctgttnc cctcccattc cagaagtaca taaaccaaag 240gctatgtctc cggaagatg 25934176DNAGlycine max 34agcataatgc cacaaatgca gaatttcaca gacgattcca tgaggatact ggaatcacat 60ttgcttccct ttaccccggt tgcattgcca caacaggcct gttcagagag cacattccct 120tgttcagaac tctgtccctc cattccagaa gtacataacc aaagggctat gtctca 17635256DNAGlycine maxunsure at all n locations 35caggaaagag acttgcacag gttgtgagtg atccacnccc taacaaaatc aggtgtttac 60tggagctgga acgcggcctc tgcttcgttt gaaaaccaat tgtcccaaga agccagcgat 120gcagataagg tcgcaaggtt tgggagatta gtgagaaact tactggtttg gcttaagtgg 180tactttggca gcttccaata tccatcttga tttagggaca tttgtcatgg agttcaataa 240catctcagaa gagttt 25636248DNAGlycine maxunsure at all n locations 36caggaaagag acttgcacag gttgtgagtg atccaagcct aacaaaatca ggtgtttact 60ggagctggaa cgcggncctg ctgcttcgtt tgaaaaccaa ttgtgcccaa gaagccagcg 120atgcagataa ggctncgcaa ggtttgggag attagtgaga aacttactgg tttgggctaa 180gtggtacttt ggcagcttcc caatatccat ctgatttagg gacattgtca ggagttcaat 240aacatctc 24837335DNAGlycine max 37ggtgtgtctc tcaaggactc caccttgttc ggtctttcat tttcagaacc tatcaaagct 60aacttcagct cttctgcatt gaggtgtcag agggaattcg aacaaaagct ctgtgctgtg 120agggccgaaa cagtggctac agcctctcca gcagttacca agtctacacc agaagggaag 180aaaacattga ggaagggcag tgttgtgata actggggctt catctggact aggcctggcc 240actgctaagg ctttggctga gacgggaaaa tggcatgtaa taatggcctg cagggattac 300ctcaaagctg caagagctgc aaaatccgct ggcat 33538258DNAGlycine max 38cggaaaatgg catgtaataa tggcctgcag ggattacctc aaagctgcaa gagctgcaaa 60atccgctggc atggctaagg aaaactacac catcatgcac taggaccttg cctcgctcga 120cagtgtccgc caatttgttg ataacttcag aagatcggaa atgccgttag atgtgctggt 180ttgcaatgct gctgtttact tgccaactgc taaggaacct accttcactg ctgagggctt 240tgaacttagt gttgggac 25839246DNAGlycine max 39aaacattgag gaagggcagt gttgtgataa ctggggcttc atctggacta ggcctggcca 60ctgctaaggc tttggctgag acgggaaaat ggcatgtaat aatggcctgc agggattacc 120tcaaagctgc aagagctgca aaatccgctg gcatggctaa ggaaaactac accatcatgc 180acttggacct tgcctcgctc gacagtgtcc gccaatttgt tgataacttc agaagatcgg 240aaatgc 24640260DNAGlycine maxunsure at all n locations 40ctgcaaganc tgcaaaatcc gctggcatgg ctaaggaaaa ctacaccatg aatgcacttg 60gaccttgcct cgctcgacag tgtccgccaa tttgttgata acttcagaag atcagaaatg 120ccgttagatg tgctggtttg ccatgctgct gtttacttgc caactgctaa ggaacctacc 180ttcactgctg agggctttga acttagtgtt gggacaaatc atctggggca tttcctcctc 240tcgcgcctgt tgcttgagga 26041278DNAGlycine maxunsure at all n locations 41attttcagaa cctatcaaag ctaacttcag ctcttctgca ttgaggttna agagggaatt 60cgaacaaaaa gctctgtgct gtgagggccg aaacagtggc tacagcctct ccagcagtta 120ccaagtctac accagaaggg aagaanacat tgaggaaggg cagtgttgtg ataactgggg 180cttcatctgg actaggcctg gccactgcta aggctttggc tgagacggga aaatggcatg 240taataatggc ctgcagggat tacctcaaag ctgcaaga 27842248DNAGlycine max 42ctgtgctgtg agggccgaaa cagtggctac agcctctcca gcagttacca agtctacacc 60agaagggaac gaaaacattg aggaagggca gtgttgtgat aactggggct tcatctggac 120taggcctggc cactgctaag gctttggctg agacgggaaa atggcatgta ataatggcct 180gcagggatta cctcaaagct gcaagagctg caaaatccgc tggcatggct aaggaaaact 240acactgtc 24843280DNAGlycine max 43gtgtctctca aggactccac cttgttcggt ctttcatttt cagaacctat caaagctaac 60ttcagctctt ctgcattgag gtgcaagagg gaattcgaac aaaagctctg tgctgtgagg 120gccgaaacag tggctacagc cttccagcag ttaccaagtc tacaccagaa gggaagaaaa 180cattgaggaa gggcagtgtt gtgataactg gggcttcatc tggactaggc ctggccactg 240ctaaggcttt ggctgagacg ggaaaatggc atgtaataat 28044269DNAGlycine max 44aaagagtggt gtgtctctca aggactccac cttgttcggt ctttcatttt cagaacctat 60caaagctaac ttcagctctt ctgcattgag gtgtaagagg gaattcgaac aaaagctctg 120tgctgtgagg gccgaaacag tggctacagc ctctccagca gttaccaagt ctacaccaga 180agggaagaaa acattgagga agggcagtgt tgtgataact ggggcttcat ctggactagg 240cctggccact gctaaggctt tggctgaga 26945236DNAGlycine max 45cgaaacagtg gctacagcct ctccagcagt taccaagtct acaccagaag ggaagcaaac 60attgaggaag ggcagtgttg tgataactgg ggcttcatct ggactaggcc tggccactgc 120taaggctttg gctgagacgg gaaaatggca tgtaataatg gcctgcaggg attacctcaa 180agctgcaaga gctgcaaaat ccgctggcat ggctaaggaa aactacacca tcatgc 23646211DNAGlycine max 46ctcgagcgtg cgagaagaga cagaaggggg aaaatggcat gtaataatgg cctgcaggga 60ttacctcaaa gctgcaagag ctgcaaaatc cgctggcatg gctaaggaaa actacaccat 120catgcacttg gaccttgcct cgctcgacag tgtccgccaa tttgttgata acttcagaag 180atcggaaatg ccgttagatg tgctggtttg c 21147276DNAGlycine

maxunsure at all n locations 47ctttttttct tcttcttgaa atggctctcc aggctgcttc tcctgttcct gcttctttct 60cggttcttaa agagggaaag agtggtgtgt ctctcaagga ctccaccttg ttcggtcttt 120cattttcaga acctatcaaa gctaacttca gctcttctgc attgaggtgc aagagggaat 180tcgancaaaa gctctgtgct gtgagggccg aaacagtggc tacagcctct ccagcagtta 240ccaagtctac accagaaggg aagnaaacat tgagga 27648269DNAGlycine max 48cttctcttgt tcctgcttct ttctcggttc ttaaagaggg aaagagtggt gtgtctctca 60aggactccac cttgttcggt ctttcatttt cagaacctat caaagctaac ttcagctctt 120ctgcattgag gtgcaagagg gaattcgaac aaaagctctg tgctgtgagg gccgaaacag 180tggctacagc ctctccagca gttaccaagt ctacaccaga agggaagaaa acattgagga 240agggcagtgt tgtgataact ggggcttca 26949279DNAGlycine max 49tagtcaaaat ctagtttcat acttttgttc ttcttcttga aatggctctc caggctgctt 60ctcttgttcc tgcttctttc tcggttctta aagagggaaa gagtggtgtg tctctcaagg 120attccacctt gttcggtctt tcattttcag aacctatcaa agctaacttc agctcttctg 180cattgaggtg caagagggaa ttcgaacaaa agctctgtgc tgtgagggcc gaaacagtgg 240ctacagcctc tccagcagtt accaagtcta caccagaag 27950257DNAGlycine max 50ttctcttgtt cctgcttctt tctcggttct taaagaggga aagagtggtg tgtctctcaa 60ggactccacc ttgttcggtc tttcattttc agaacctatc aaagctaact tcagctcttc 120tgcattgagg ttcaagaggg aattcgaaca aaagctctgt gctgtgaggg ccgaaacagt 180ggctacagcc tctccagcag ttaccaagtc tacaccagaa gggaagataa cattgaggaa 240gggcagtgtt gtgataa 25751243DNAGlycine max 51ggctgcttct cttgttcctg cttctttctc ggttcttaaa gagggaaaga gtggtgtgtc 60tctcaaggac tccaccttgt tcggtctttc attttcagaa cctatcaaag ctaacttcag 120ctcttctgca ttgaggtgca agagggaatt cgaacaaaag ctctgtgctg tgagggccga 180aacagtggct acagcctctc cagcagttac caagtctaca ccagaaggga agaaaacatt 240gag 24352277DNAGlycine maxunsure at all n locations 52caatattgta aaactcaaaa tctagtttca tacttttttt cttcttcttg aaatggctct 60ccaggctgct tctcttgttc ctgcttcttt ctcggttctt aaagagggaa agagtggtgt 120gtctctcaag gactccacct tgttcggtct ttcattttca gaacctatca aagctaactt 180cagctcttct gcattgaggt ncaagaggga attcgaacaa aagctctntg ctgtgagggc 240cgaaacagtg gctacagcct ctccagcagt taccaag 27753271DNAGlycine maxunsure at all n locations 53ctttttttct tcttcttgaa tggctctcca ggctgcttct cttgancctg cttccttctc 60ggttcttaaa gagggaaaga gtggtgtgtc tctcaaggac tccaccttgt tcggtctttc 120attttcagaa cctatcaaag ctaacttcag ctcttctgca ttgaggttaa gagggaattc 180gaacaaaagc tcngtgctgt gagggccgaa acagtggcta cagcctctcc agcagttacc 240aagtctacac cagaaggcaa nnaacattga g 27154269DNAGlycine maxunsure at all n locations 54cnatattgta aaactcaaaa tctagtttca tacttttttt cttcttcttg aaatggctct 60ccaggctgct tctcttgttc ctgcttcttt ctcggttctt aaagagggaa agagtggtgt 120gtctctcaag gactccacct tgttcggtct ttcattttca gaacctatca aagctaactt 180cagctcttct gcattgaggt ccaagaggga attcgaacaa aagctctgtg ctgtgagggc 240cgaaacagtg gctanagcct ctccagcag 26955282DNAGlycine max 55tcaaaatcta gtttcatact tttcatcttc ttcttgaaat ggctctccag gctgcttctc 60ttgttcctga ttctttctcg gttcttaaag acggtgagat gtggtgtgtc tctcaaggac 120tccacctagt tcggtctggc attttcagaa cctatcaaag ctaacttaag ctcttctgca 180ttgaggtgca agagggattc cgcacaaaag ctctgtgctg tgagtgccga gacagtggct 240acagcgtctg cagcagttac caagtctaca cgagaaggga ag 28256263DNAGlycine max 56acttctcttg ttcctgcttc tttctcggtt cttaaagagg gacagagtgg tgtgtctctc 60aaggactccg cttgttcggt ctttcatttt cagaacctat caaagctaac ttcagctctt 120ctgcattgag gtgcaagagg gaattcgaac aatcgctctg tgctgtgagg gccgaaacag 180tggttacagc ctctccagca gttaccaagt ctacaccaga tgggaagaaa acattgagtg 240aaggagtgtg gtgaaactgg ggc 26357313DNAGlycine max 57gaaatggctc tccaggctgc ttctcttgtt cctgcttctt tctcggttct taaagaggga 60aagagtggtg tgtctctcaa ggactccacc ttgttcggtc tttcattttc agaacctatc 120aaagctaact tcagctcttc tgcattgagg tgcaagaggg aattcgaaca aaagctctgt 180gctgtgaggg ccgaaacagt ggctacagcc tctccagcag ttaccaagtc tacaccagaa 240ggcaagaaaa cattgaggaa gggcagtgtt gtgataactg gggcttcatc tggacgaggc 300ctggccactg cta 31358266DNAGlycine max 58ccgtgataac acactaacac caccacttca tcaactttac ttgacaacaa tattgtaaaa 60ctcaaaatct agtttcatac ttttgttctt cttcttgaaa tggctctcca ggctgcttct 120cttgttcctg cttctttctc ggttcttaaa gagggaaaga gtggtgtgtc tctcaaggac 180tccaccttgt tcggtctttc attttcagaa cctatcaaag ctaacttcag ctcttctgca 240ttgaggtgca agagggaatt cgaaca 26659277DNAGlycine max 59caccatcact tcatcaactt tacttgacaa caatattgta aaactcaaaa tctagtttca 60tacttttttt cttcttcttg aaatggctct ccaggctgct tctcttgttc ctgcttcttt 120ctcggttctt aaagagggaa agagtggtgt gtctctcaag gactccacct tgttcggtct 180ttcattttca gaacctatca aagctaactt cagctcttct gcattgaggt gcaagaggga 240attcgaacaa aagctctgtg ctgtgagggc cgaaaca 27760151DNAGlycine max 60gcatctttct cggttcttaa agagggaaag actggtgtgt cactcacgga ttccaccttg 60tacggtcttt cattttcaga acctatcaaa gctaacttca gctcttctgc attgaggtgc 120aagagggaat tcgaacaaaa actctgtgct g 15161266DNAGlycine max 61caccacttca tcaactttac ttgacaacaa tattgtaaaa ctcaaaatct agtttcatac 60tttttttact cttcttgaaa tggctctcca ggctgcttct cttgttcctg cttctttctc 120ggttcttaaa gagggaaaga gtggtgtgtc tctcaaggac tccaccttgt tcggtctttc 180attttcagaa cctatcaaag ctaacttcag ctcttctgca ttgaggtgca agagggaatt 240cgaacaaaag ctctgtgctg tgaggg 26662229DNAGlycine max 62ttcatcaact ttacttgaca acaatattgt aaaactcaaa atctagtttc atactttttt 60tcttcttctt gaaatggctc tccaggctgc ttctcttgtt cctgcttctt tctcggttct 120taaagaggga aagagtggtg tgtctctcaa ggactccacc ttgttcggtc tttcattttc 180agaacctatc aaagctaact tcagctcttc tgcattgagg tgcaagagg 22963268DNAGlycine max 63cccgtgataa cacactaaca ccatcacttc atcaacttta cttgacaaca atattgtaaa 60actcaaaatc tagtttcata cttttattcg tcttctttaa atggctctcc aggctgcttc 120tcttgttcct gcttctttct cggttcttaa atagggaaag agtggtgtgt ctctcaagga 180ctccaccttg ttcggtcttt cattttcaga acctatcaaa gctaacttca gctcttctgc 240attgaggttc aagagggaat tcgaacaa 26864278DNAGlycine maxunsure at all n locations 64tatnatacca cttcatcaac ctnacnctga caacaatatt gtaaaactcn naatctagtt 60tcatactttt tttcttcttc ttgaaatggc tctccaggct gcttctcttg ttcctgcttc 120tttctcggtt cttaaagagg gaaagagtgg tgtgtctctc aaggactcca ccttgttcgg 180tctttcattt tcagaaccta tcaaagctaa cttcagctct tctgcattga ggtntcaaga 240gggaattcga acaaaagctc tgtgctgtga gggccgaa 27865275DNAGlycine max 65ttcatcaact ttacttgaca acaatattgt aaaattcaaa atctagtttc atacttttat 60tcttcttctt gaaatggctc tccaggctgc ttctcttgtt cctgcttctt tctcggttct 120taaagaggga aagagtggtg tgtctctcaa ggactccacc ttgttcggtc tttcattttc 180agaacctatc aaagctaact tcagctcttc tgcattgagg tttaagaggg aattcgaaca 240aaagctctgt gctgtgaggg ccgaaacagt ggcta 27566344DNAGlycine maxunsure at all n locations 66caatattgta naactcaaaa tctagtttca tacttttctt ctacttcttg aaatggctct 60ccaggctgct tctcttgttc ctgcttcttt ctcggttctt aaagagggaa agagtggtgt 120gtttctcaag gactccacct tgttcggtct ttcattttca gaacctttta tagctaactt 180cagctcttct gcattgaggt gtaagaggga attcgaacaa aagctctgtg ctgtgagggc 240cgaaacagtg gctacagcct ctccagcagt taccaagtct acaccagaag ggacgtcaac 300attgaggaag ggcagtgttg tgataactgg ggcttcatct ggac 34467255DNAGlycine max 67cgccgtgata acacactaac accaccactt catcaacttt acttgacaac aatattgtaa 60aactcaaaat ctagtttcat actttttttc ttcttcttga aatggctctc caggctgctt 120ctcttgttcc tgattcttac tcggttctta aagagggaaa gagtggtgtg tctctcaagg 180actccacctt gttcggtctt tcattttcag aacctatcaa agctaacttc agctcttctg 240cattgaggtg caaga 25568249DNAGlycine max 68ttttcattac cgccgtgata acacactaac accaccactt catcaacttt acttgacaac 60aatattgtaa aactcaaaat ctagtttcat actttttttc ttcttcttga aatggctctc 120caggctgctt ctcttgttcc tgcttctttc tcggttctta aagagggaaa gagtggtgtg 180tctctcaagg actccacctt gttcggtctt tcattttcag aacctatcaa agctaacttc 240agctcttct 24969249DNAGlycine max 69cacactaaca ccaccacttc atcaacttta cttgacaaca atattgtaaa actcaaaatc 60tagtttcata ctttttttct tcttcttgaa atggctctcc aggctgcttc tcttgttcct 120gcttctttct cggttcttaa agagggaaag agtggtgtgt ctctcaagga ctccaccttg 180ttcggtcttt cattttcaga acctatcaaa gctaacttca gctcttctgc attgaggttc 240aagagggaa 24970294DNAGlycine max 70caatattgta aaactcaaaa tctagtttca tacttttttt cttcttcttg aaatggctct 60ccaggctgct tctcttgttc ctgcttcttt ctcggttctt aaagagggaa agagtggtgt 120gtctctcaag gactccacct tgttcggtct ttcattttca gaacctatca aagctaactt 180cagctcttct gcattgaggt gcaagaggga attcgaacaa aagctctgtg ctgtgagggc 240cgaaacagtg gctacagcct ctccagcagt taccaagtct acaccagaag ggaa 29471270DNAGlycine max 71ctccaggctg cttctcttgt tcctgcttct ttctcggttc ttaaagaggg aaagagtggt 60gtgtctctca aggactccac cttgttcggt ctttcatttt cagaacctat caaagctaac 120ttcagctctt ctgcattgag gtgcaagagg gaattcgaac aaaagctctg tgctgtgagg 180gccgaaacag tggctacagc ctctccagca gttaccaagt ctacaccaga aggcaagata 240acattgagaa gggcagtgtt gtgataactg 27072254DNAGlycine max 72attaccgccg tgataacaca ctaacaccac cacttcatca actttacttg acaacaatat 60tgtaaaactc aaaatctagt ttcatacttt ttttcttctt cttgaaaggc tctccaggct 120gcttctcttg ttcctgcttc tttctcggtt cttaaagagg gaaagagtgg tgtgtctctc 180aaggactcca ccttgttcgg tctttcattt tcagaaccta agctaacttc agctcttctg 240cattgaggtg caag 25473100DNAGlycine maxunsure at all n locations 73ccctgcaggc cattattaca aagctgcaag agctgcaaaa tccgctggca tggctaagga 60aaactacacc atcatgcanc ttggaccttg cctcgctcga 10074262DNAGlycine max 74cgccgtgata acacactaac accaccactt catcaacttt acttgacaac aatattgtaa 60aactcaaaat ctagtttcat actttttttc ttcttcttga aatggctctc caggctgctt 120ctcttgttcc gcttctttct cggttcttaa agagggaaag agtggtgtgt ctctcaagga 180ctccaccttg ttcggtcttt cattttcaga acctatcaaa gctaacttca tcttctgcat 240tgaggtgcaa gagggaattc ga 26275184DNAGlycine max 75gtgataacac actaacacca ccacttcatc aactttactt gacaacaata ttgtaaaact 60caaaatctag tttcatactt tttttcttct tcttgaaatg gctctccagg ctgcttctct 120tgttcctgct tctttctcgg ttcttaaaga gggaaagagt ggtgtgtctc tcaaggactc 180cacc 18476229DNAGlycine max 76ggaaccacac atttttcatt accgccgtga taacacacta acaccaccac ttcatcaact 60ttacttgaca acaatattgt aaaactcaaa atctggtttc atactttttt tcttcttctt 120gaaatggctc tccaggctgc ttctcttgtt cctgcttctt tctcggttct taaagaggga 180aagagtggtg tgtctctcaa ggactccacc ttgttcggtc tttcatttt 22977270DNAGlycine maxunsure at all n locations 77attaccgtcg tgataacaca ctaacaccac cacttcatca actttacttg acaacaatat 60tgtaaaactc aaaatctagt nnnnnnnnnn nnnnnnnnnn nnngaaatgg ctctccaggc 120tgcttctctt gttcctgctt ctttctcggt tcttaaagag ggaaagagtg gtgtgtctct 180caaggactcc accttgttcg gtctttcatt ttcagaacct atcanagcta acttcagctc 240ttctgcatga gngntagang gantcgaaca 27078267DNAGlycine max 78ggctgcgaga agacgacaga aggggaacca cacatttttc attaccgccg tgataacaca 60ctaacaccac cacttcatca actttacttg acaacaatat tgtaaaactc aaaatctagt 120ttcatacttt ttttcttctt cttgaaatgg ctctccaggc tgcttctctt gttcctgctt 180ctttctcggt tcttaaagag ggaaagagtg gtgtgtctct caaggactcc accttgttcg 240gtctttcatt ttcagaacct atcaaag 26779158DNAGlycine max 79tcaaaatcta gtttcatact ttttttcttc ttcttgaaat ggctctccag gctgcttctc 60ttgttcctgc ttctttctcg gttcttaaag agggaaagag tggtgtgtct ctcaaggact 120ccaccttgtt cggtctttca ttttcagaac ctatcaaa 15880278DNAGlycine max 80cacactaaca ccaccacttc atcaacttta cttgacaaca atattgtaaa actcaaaatc 60tagtttcata ctttttttct tcttcttgaa atggctctcc aggctgcttc tcttgttcct 120gcttctttct cggttcttaa gagggaaaga gtggtgtgtc tctcaaggac tccaccttgt 180tcggtctttc attttcagaa cctatcaaag ctaacttcag ctcttctgca ttgaggtgca 240agagggaatt cgaacaaaag ctctgtgctg tgagggcc 27881285DNAGlycine max 81cacggctgcg aaagacgaca gaaggggacc acacattttt cattaccgcc gtgataacac 60actaacacca ccagctcatc aactttactt gacaacaata ttgtaaaact caaaatctag 120tttcatactt tttttcttct tcttgaaatg gctctccagg ctgcttctct tgttcctgct 180tctttctcgg ttcttaaaga gggaaagagt ggtgtgtctc tcaaggactc caccttgttc 240ggtctttcat tttcagaact atcaaagcta attcagctct tctgc 28582269DNAGlycine max 82ggttaccatt atttctttat aactatacta ctcatcagct gcatggtatt tttgctttca 60ttgttggtgt tgttgttgat ccacttcatc aactttactt gacaacaaga ttgtaaaact 120caaaatctag tttcatactt tttttcttct tcttgaaatg gctctccagg ctgcttctct 180tgttcctgct tctttctcgg ttcttaaagc gggcaagagt ggtgtgtctc tcaaggactc 240caccttgttc ggtctttcat tttcagaac 26983260DNAGlycine max 83acggcgagaa gacgacagaa ggggaaccac acatttttca ttaccgccgt gataacacac 60taacaccacc acttcatcaa ctttacttga caacaatatt gtaaaactca aaatctagtt 120tcatactttt tttcttcttc ttgaaatggc tctccaggct gcttctcttg ttcctgcttc 180tttctcggtt cttaaagagg gaaagagtgg tgtgtctctc aaggactcca ccttgttcgg 240tctttcattt tcagaaccta 26084108DNAGlycine max 84ttcagctctg ctgcattgag gtgccagagg gaattcgaac aaaagctctg tgctgtgagg 60gccgaaacag tggctacagc ctctccagca gttaccaagt ctacacca 10885258DNAGlycine max 85caatattgta aaactcaaaa tctagtttca tacttttttt cttcttcttg aaatggctct 60ccaggctgcc tctcttgttc ctgcttcttt ctcggttctt aaagagggaa agagtggtgt 120gtctctcaag gactcacctt gttcggtctt tcattttcag aacctatcaa agctaacttc 180agctcttctg cattgaggtg taagagggaa ttcgaacaaa agctctgtgc tgtgagggcc 240gaaacagtgg ctacagcc 25886250DNAGlycine max 86caatattgta aaactcaaaa tctagtttca tacttttttt cttcttcttg aaatggctct 60ccaggctgct tctcttgttc ctgcttcttt ctcggttctt aaagagggaa agagtggtgt 120gtctctcaag gctccacctt gttcggtctt tcattttcag aacctatcaa agctaacttc 180agctcttctg cattgaggtg caagagggaa ttcgaacaaa agctctgtgc tgtgaggcga 240aacagtggct 25087260DNAGlycine maxunsure at all n locations 87caaaaatttg gccctttgag ggttcagtca gtggcaacaa caactccagg agtcaccaag 60gcttcaccag aaggcaagaa nactttgagg aaaggcagtg ttattatcac tggggcttcc 120tctggattag gcctggccac tgctaaggct ttggctgaga caggaaagtg gcatgtgata 180atggcctgcc gggatttcct caaagccgaa anngctgcga aatctgccgg cattgctaag 240gaaaactaca ctattatgca 26088281DNAGlycine max 88caacaaaaaa ttggcccttt gagggttcag tcagtggcaa caaccactcc aggagtcacc 60aaggcttcac cagaaggcaa gaaaactttg aggaaaggca gtgttattgt cactgggctt 120cctctggatt aggcctggcc acggccaagg ctttggctga gacaggaaag tggcatgtga 180ttatgcactg cagggatttc ctcaaagctg agagggctgc aaaatctgct ggcattgcta 240aggaaattgt gtctcttgat agtgtgaggc aatttgtgga t 28189385DNAGlycine max 89ctttgaactt agtgttgggc caaataattt gggcgttttc gtctctctcg cctgttgctt 60gaggacttgg aaaaatccga ttacccttca aagcgcttga tcatcgttgg ttcaatatca 120cggaacacac acacattggc tggtaatgta cctcccaagg ctaaccttgg tgacttgagg 180ggacttcaag gtggtttgaa tgggcttaac agctcagcca tgattgatgg tggagacttc 240gatggtgcca aggcgtacaa ggacagcaaa gtctgcaata tgctcacaat gcaagaattc 300cacagacgat ttcatgagga aaactgaatc acatttgctt tcctttaacc ccggtgcatt 360gccacaacag gcctgttcag agagc 38590241DNAGlycine maxunsure at all n locations 90gataacttca gaagatcgga aatgccgtta gatgtgctgg tttgcaatgc tgctgtttac 60ttgccaactg ctaaggaacc taccttcact gctgagggct ttgaacttag tgttgggaca 120aatcatctgg ggcatttcct cctctcgcgc ctgttgcttg aggacttgga aaaatccgat 180tacccttcaa agcgcttgat catcgttggt tcaataacag ggnacacaaa cacattggct 240g 24191267DNAGlycine max 91ctcctctcgc gcctgttgct tgaggacttg gaaaaatccg attacccttc aaagcgcttg 60atcatcgttg gttcaataac agggaacaca aacacattgg ctggtaatgt acctcccaag 120gctaaccttg gtgacttgag gggacttcag ggtggtttga atgggctaaa cagctcagcc 180atgattgatg gtggagagat cgatggtgcc aaggcgtaca aggacagcaa agtctgcaat 240atgctcacaa tgcaagaatt ccacaga 26792256DNAGlycine max 92ttagatgtgc tggtttgcaa tgctgctgtt tacttgccaa ctgctaagga acctaccttc 60actgctgagg gctttgaact tagtgttggg acaaatcatc tggggcattt cctcctctcg 120cgcctgttgc ttgaggactt ggaaaaatcc gattaccctt caaagcgctt gatcatcgtt 180ggttcaataa cagggaacac aaacacattg gctggtaatg tacctcccaa ggctaacctt 240ggtgacttga ggggat 25693260DNAGlycine max 93cttcactgct gagggctttg aacttagtgt tgggacaaat catctggggc atttcctcct 60ctcgcgcctg ttgcttgagg acttggaaaa atccgattac ccttcaaagc gcttgatcat 120cgttggttca ataacaggga acacaaacac attggctggt aatgtacctc ccaaggctaa 180ccttggtgac ttgaggggac ttcagggtgg tttgaatggg ctaaacagct cagccatgat 240tgatggtgga gattcgatgg 26094274DNAGlycine maxunsure at all n locations 94cntaccttca ctgctgaggg ctttganctt antgttngng acaaattcat ctggggcatt 60tcctcctctc gcgcctgttg cttgaggact tggaaaaatc cgattaccct tcaaagcgct 120tgatcatcgt tggttcaata acagggaaca caaacacatt ggctggtaat gtactcccaa 180ggctaacctt ggtgacttga ggggacttca gggtggtttg aatgggctaa acagctcagc 240catgattgat ggtggagatt cgatggtgcc aagc

27495284DNAGlycine max 95cagtattgtg aaatgttgaa agcagacgag tggcctgttt gtgcatttat ttctcaagat 60tgtcgtccag caaatccatc ggaagaagcg cacaatgttc aaacatcgta tgaagtgtgg 120gagaagacat tagagatgat tggccttccc tcagatgctg tggaaaggct tttagatggg 180gaagaagtta aatgccgtta tggacaagaa cagtaatcta atatacaata tctcccttaa 240tctgtaaggg cacttccatt atttatagct agtaatgagc attt 28496265DNAGlycine maxunsure at all n locations 96aagagagaga tggcaacgac gacgtcgtct tcaagcgagg nagcaccgaa cactaagaag 60aacaagaagg agcgtttagg ttggntagaa tggttaagag gttggttcta tttggtctac 120gaaatgctct ttcagcgcat catggcgagc cacttgcaca accctatgcc tctccctcct 180gtaaacgacc tcacttgcat tgtcaccggc tccaccagcg gcattggcct cgaaattgct 240aggcaattgg ctcagtcagg ggccc 26597135DNAGlycine maxunsure at all n locations 97ggaaagaaca atggttggca gtaggtatac tacaagtaac tcctcaatcc catgtaagan 60aacaaaaggc agcttcttta atgccagtat tgcacaacac ctcagactag tacaanaaaa 120aacaaagaaa agggg 13598129DNAGlycine max 98ccatttgcca ttggatggcg ctgctagaat ttgtactggt gccaccagtt tcctctccct 60ttatgtccca gatgagtacc caagtggcaa aaattagatt agactaatat atatatattg 120ttttatcag 12999270DNAGlycine max 99gtccaggccc ggtggcggcg gtggcattag cagggtcctt caagacggtg ccgtttggga 60aaaaggctgg ggttaatgcc cctgttgttt acggtgtcat gccacctgac gcatatcgtg 120ctgccaaggg tgttcctacc gatcaaaaac ctggtcctgt gcctttcttc gctgctggaa 180tcagctccgt tttacaccca aagaacccgt ttgcccctac cctacatttc aactatcgct 240attttgaaac cgatgctcct aaagatgctc 270100264DNAGlycine maxunsure at all n locations 100aattgcgaag gggacgatat gttgaattca atttggtata tgatagnggt acaacatttg 60gnctgaaaac tggagggaga atagagagta tacttgtttc tctcccactg actgctcggt 120gggaatacga tcataaaccg gaagaaggaa gcgaagaatg gaaactcttg gacgcatgca 180tcaaccccaa ggaatggatc taattcatca gttgaccccc caatttgtca gctttttaat 240ttaataataa gggagcttgt ttct 264101249DNAGlycine max 101ctcccttatt attaaattaa aaagctgaca aattgggggg tcaactgatg aattagatcc 60attccttggg gttgatgcat gcgtccaaga gtttccattc ttcgcttcct tcttccggtt 120tatgatcgta ttcccaccga gcagtcagtg ggagagaaac aagtatactc tctattctcc 180ctccagtttt cagtccaaat gttgtacccc tatcatatac caaattgaat tcaacatatc 240gtccccttc 249102262DNAGlycine max 102ggagatgctc ctttcctttg ctactgaatg tgcaaattct gttattcctg cttatttacc 60tatcatagag aaaaggaagg atttgccctt caatgatcat cagaaagcat ggcaacaatt 120gcgaagggga cgatatgttg aattcaattt ggtatatgat aggggtacaa catttggact 180gaaaactgga gggagaatag agagtatact tgtttctctc ccactgactg ctcggtggga 240atacgatcaa aaccggaaga ag 262103240DNAGlycine max 103agatgctcct ttcctttgct actgaatgtg caaattctgt tattcctgct tatttaccta 60tcatagagaa aaggaaggat ttgcccttca atgatcatca gaaagcatgg caacaattgc 120gaaggggacg atatgttgaa ttcaatttgg tatatgatag gggtacaaca tttggactga 180aaactggagg gagaatagag agtatacttg tttctctccc actgactgct cggtgggaat 240104249DNAGlycine max 104acggctgcga gaagacgaca gaaggggatg atcttaatga ctatgatcag gagatgctcc 60tttcctttgc tactgaatgt gcaaattctg ttattcctgc ttatttacct atcatagaga 120aaaggaagga tttgcccttc aatgatcatc agaaagcatg gcaacatttg cgaacgggga 180cgatatgttg aattcaattt ggtatatgat aggggtacaa catttggact gaaaactgga 240gggagaata 249105250DNAGlycine maxunsure at all n locations 105aattgcgnag gggangatat gntgaatnca attnggtana tgntannggt acaacanttg 60gactgaatnc tggaggggag aatagagagt atacttgttt ctctcncact gactgctcgg 120tgggaatacg atcatnaacc ggnagangga agcgaagact ggnaactctt ggncgcatgc 180atnaacccca aggaatggat ctaattcatc agttgacccc ccaatttgtc agctttttaa 240tttaataata 250106268DNAGlycine max 106ggatttgccc ttcaatgatc atcagaaagc atggcaacaa ttgcgaaggg gacgatatgt 60tgaattcaat ttggtatatg ataggggtac aacatttgga ctgaaaactg gagggagaat 120agagagtata cttgtttctc tcccactgac tgctcggtgg gaatacgatc ataaaccgga 180agaaggaagc gaagaatgga aactcttgga cgcatgcatc aaccccaagg aatggatcta 240attcatcagt tgacccccca atttgtca 268107268DNAGlycine max 107acggctgcga gaagacgaca gaaggggaga aaaggaagga tttgcccttc aatgatcatc 60agaaagcatg gcaacaattg cgaaggggac gatatgttga attcaatttg gtatatgata 120ggggtacaac atttggactg aaaactggag ggagaataga gagtatactt gtttctctcc 180cactgactgc tcggtgggaa tacgatcata aaccggaaga aggaagcgaa gaatggaaac 240tcttggacgc atgcatcaac cccaagga 268108321DNAGlycine max 108ggaagacctt atcatctccg aatttcattt tcagaagcct ctttgggaat caaatccgaa 60gcatgatgca ttgtgcgagc attgtctcgg ctccgtccta cgcgttccct tttctctctg 120gctccgcttc cactactcca actgcgatct cgctcactaa gcgcagttgg aagccacctc 180cgagcatggc aaaaggccca gtcagagcca ccgtttctat agagaaagag accccggagg 240ccaatcgtcc cgaaacgttt ctcagaggag tggacgaggc ccagtcttcc acttcggttc 300gggcccgctt cgagaagatg a 321109282DNAGlycine max 109cacatccgaa gcatgatgca ttgtgcgagc attgtctcgg ctccgtccta cgcgttccct 60tttctctctg gctccgcttc cactactcca actgcgatct cgctcactaa gcgcagttgg 120aagccacctc cgagcatggc aaaaggccca gtcagagcca ccgtttctat agagaaagag 180accccggagg ccaatcgtcc cgaaacgttt ctcagaggag tggacgaggc ccagtcttcc 240acttcggttc gggcccgctc tcgagaagat gataagggac gc 282110260DNAGlycine max 110ccttatcatc tccgaatttc attttcagaa gcctctttgg gaatcaaatc cgaagcatga 60tgcattgtgc gagcattgtc tcggctccgt cctacgcgtt cccttttctc tctggctccg 120cttccactac tccaactgcg atctcgctca ctaagcgcag ttggaagcca cctccgagca 180tggcaaaagg cccagtcaga gccaccgttt ctatagagaa agagaccccg gaggccaatc 240gtcccgaaac gtttctcaga 260111269DNAGlycine max 111ctctttggga atcaaatccg aagcatgatg cattgtgcga gcattgtctc ggctccgtcc 60tacgcgttcc cttttctctc tggctccgct tccactactc caactgcgat ctcgctcact 120aagcgcagtt ggaagccacc tccgagcatg gcaaaaggcc cagtcagagc cacgtttcta 180tagagaaaga taccccggag gccaatcgtc ccgaaacgtt tctcagagga gtggacgagg 240cccagtcttc cacttcggtt cgggcccgc 269112260DNAGlycine max 112tgtgcgagca ttgtctcggc tccgtcctac gcgttccctt ttctctctgg ctccgcttcc 60actactccaa ctgcgctctc gctcactaag cgcagttgga agccacctcc gagcatggca 120aaaggcccag tcagagccac cgtttctata gagaaagaga ccccggaggc caatcgtccc 180gaaacgtttc tcagaggagt ggacgaggcc cagtcttcca cttcggttcg ggcccgcttc 240gagaagatga taagggaggc 260113279DNAGlycine maxunsure at all n locations 113gaagacttta tcatttccga atttcntttt cagangcctc tttgggaatc anntccnnng 60catgatgcat tgtngcgagc nttgtctacg gctccgtcct acgcgttccc ttttcgctct 120ggctccgctt ccactactcc aactgcgntc tcgctcacta agcgcagttg gaagccacct 180ccgagnatgg caaaaggccc agtcagagcc accgtttcta tagagaaaga gaccccggag 240gccaatcgtc ccgaaacgtt tctcagagga gtggacgag 279114247DNAGlycine max 114ctccgaattt cattttcaga agcctctttg ggaatcaaat tggagtgtct gcaatccact 60ccgaagcatg atgcattgtg cgagcattgt ctcggctccg tcctacgcgt tcccttttcg 120ctctggctcc gctctccact actccaactg cgatctcgct ctctaagcgc agttggaagc 180cacctccgag catggcaaaa gcccagtcag agccaccgtt tctatagaga aagagacccc 240ggaggcc 247115253DNAGlycine max 115cagaagcctc tttgggaatc aaatccgaag catgatgcat tgtgcgagca ttgtctcggc 60tccgtcctac gcgttccctt ttctctctgg ctccgcttcc actactccaa ctgccctctc 120gctcactacg cgcagttgga agccacctcc gagcatggca aaaggcccag tcagagccac 180cgtttctata gagatagaga ccccggaggc caatcgtccc gaaacgtttc tcagaggagt 240ggacgaggcc cag 253116268DNAGlycine max 116tcgagcgcgt tcccttttct ctctggctcc gcttccacta ctccacatgc gctctcgctc 60actaagcgca gttggaagcc acctccgagc atggcaaaag gcccagtcag agccaccgtt 120tctatagaga aagagacccc ggaggccaat cgtcccgaaa cgtttctcag aggagtcgtc 180gaggcccagt cttccacttc ggttcgggcc cgcttcgaga agatgataag ggaggcccag 240gacaccgtgt gcagtgccct cgaggccg 268117238DNAGlycine max 117atccgaagca tgatgcattg tgcgagcatt gtctcggctc cgtcctacgc gttccctttt 60ctctctggct ccgcttccac tactccaact gcgatctcgc tcactaagcg cagttggaag 120ccacctccga gcatggcaaa aggcccagtc agagccaccg tttctataga gaaagacacc 180ccggaggcca atggtcccga aacgtttctc agaggagtgg acgaggccca ttcttcca 238118250DNAGlycine max 118tccgaagcat gatgcattgt gcgagcattg tctcggctcc gtcctacgcg ttcccttttc 60tctctggctc cgcttccact actccaactg ccctctcgct cactaagcgc agttggaagc 120cacctccgag catggcaaaa ggaccagtca gagccaccgt ttctacagag acagagaccc 180cggaggccaa tcgtcccgaa acgtttctca gaggagtgga cgaggccaag tcttccactt 240cggttcgggc 250119267DNAGlycine max 119actcgagccg attcggctcg agctctttgg gaatcaaatc cgaaacatga tgcattgtgc 60gaccattgtc tcggctccgt cactacgcgt tcccttttct ctctggctcc gcttccacta 120ctccaactac tactctcgct cactaagcgc agttggaagc cacctccgag catggcaaaa 180ggcccagtca gagccaccgt ttctatagag acagacaccc cggaagccaa ttctcccgaa 240acgtttctca gacgactgga cgaggcc 267120119DNAGlycine max 120tcattttcag aagcctcttt gggaatcaaa tccgaagcat gatgcattac gcgagcattg 60tctcggctcc gtcctacgcg ttcccttttc tctctggctc cgcttccaca caacatacg 119121117DNAGlycine maxunsure at all n locations 121cgaatttcat tttcagaagc ctctttggga atcaaatccg aagcatgatg cattgngcga 60gcattgtctc ggctccgtcc tacgcgttcc cttttctctc tggctccgct tccacaa 11712294DNAGlycine max 122caaatccgaa gcatgatgca ttgtgcgagc attgtctcgg ctccgtccta cgcgttccct 60tttctctctg gctccgcttc cacacaacat acga 9412381DNAGlycine max 123cattttcaga agcctctttg ggaatcaaat ccgaagcatg atgcattgtg cgagcattgt 60ctcggctccg tcctacgcgt t 81124246DNAGlycine maxunsure at all n locations 124cgagacccgg aggccaatcg tcncgaaacg tttctcagag gagtggacga gtgccagtct 60tccacttcgg ttcgggcntc gttcgagaag atgataaagg gaggcccagg acaccgtgtg 120cagtgccctc gaggccgctg atggtggggc ccagttcaag gaggacgttt ggtccaggcc 180cggtggcggc ggtggcatta gcagggtcct tcaagacggt gccgtttggg agaaggctgg 240ggttaa 246125261DNAGlycine max 125gaaagagacc ccggaggcca atcgtcccga aacgtttctc agaggagtgg acgaggccca 60gtcttccact tcggttcggg cctgcttcga gaagatgata agggaggccc aggacaccgt 120gtgcagtgcc ctcgaggccg ctgatggtgg ggcccagttc atggaggacg tttggtccag 180gcccggtggc ggcggtggca ttagcagggt ccttcaagac ggtgccgttt gggagaaggc 240tggggttaat gtctctgttg t 261126239DNAGlycine maxunsure at all n locations 126accaatcgtc ccgaaacgtt tctcagagga gtggacgagg cccagtcttc cacttcggtt 60cgggcccgct tcgagaagat gataagggag gcccaggaca ccgtgtgcag tgccctcgag 120gccgctgatg gtggggccca gttcaaggag gacgtttggt ccaggcccgg tggcggcggt 180ggcnncagca ggtccttcaa gacggtgccg tttgggagaa ggctggggtt aatgtctct 239127162DNAGlycine max 127atcaagtgct tgttatgatg agtcagaatg ttagcttgtt gtactaggtg gattgtaaat 60cacgtatttt gctagagtca tccgcgtaaa gcgtgaaaat gcagaaaatt acaaatgtct 120aggctgcgtc tgtagtatac ctactgccaa ccattgttct tt 162128114DNAGlycine maxunsure at all n locations 128atcaagtgct tgttcatgat ggtcagaatg ttagcttgtt gtactaggtg gattgtaaat 60cacgtatctt gctagagtnc tccgcgcgga gcgtgaanat gcagagaatt acaa 114129253DNAGlycine max 129ggcgtctgcc aaaaccaaaa ggtcagactg ttggatcttt ccggaaggga cttaccatgt 60tgcctgatgc aatttctgcc agactaggca acaaagtaaa gttatcttgg aagctttcaa 120gtattagtaa actggatagt ggagagtaca gtttgacata tgaaacacca gaaggagtgg 180tttctttgca gtgcaaaact gttgtcctga ccattccttc ctatgttgct agtacatgcc 240tgcgtcctct gtc 253130298DNAGlycine maxunsure at all n locations 130gctgcagatg cactttcaaa gttttattac cctccagttg ctgcagtttc catatcctat 60ccanaagaag ctattagatc agaatgcttg atagatggtg agttgaaggg ggttggtcaa 120ttgcatccac gtagacaagg agtggaaaca ttaggaacta tatacagctc atcactattc 180cccaaccgag caccacgacg gaaggttcta ctcttgaatt acattggagg agcaactaat 240actggaattt tatcgaagac ggacagtgaa cttgtggaaa cagttgatcg agatttga 298131283DNAGlycine max 131caattatata taatctcctg ctgactcgtc tttttctttg gaataatgat atactgtcaa 60aaaccatata taatctcctg ctgacacatc tttttctttt cttttcttta tatcattttc 120cttattagtt tctttgttta ctgcagtgac gagcttagga aaattgttac ttctgacctg 180agaaagttgt tgggagcaga gggggaacca acatttgtta accatttcta ttggagtaaa 240ggctttcctt tgtatggacg taactatggg tcagttctta agc 283132250DNAGlycine max 132tgacaatttt gatgatagag gtggataata aagctgcagt ccttggttat atcggggcac 60cgctcactct ggcatcacat gtgattgaag gtggttcatc accaaacttc tcgcaaataa 120agagattggc tttctcagca tccaagatcc tgcactcgtt actgcagaag tttacgacat 180ctctggcgag atacattctc taccaagctg acaatggagc tcaagctgtt cagatctttg 240attcatgggc 250133235DNAGlycine max 133tgacaatttt gaggaaagag gtggataata aagctgcagt ccttggtttt gtcggggcac 60cgttcactct ggcatcatat gtggttgaag gtggttcatc aaaaaacttc tcaaaaataa 120agagattggc tttctcagaa tccaagatcc tgcactcgtt actgcagaag tttacaacat 180caatggcaag atacattcaa taccaagctg acaatggagc tcaagctgtt cagat 235134282DNAGlycine maxunsure at all n locations 134gtggacaact accacctgaa atgtgggaac gctggtcaaa gccttatatc aaagagattg 60taaatttggt cangaaaaaa tgccctgggg taccaattgt tctttatata aacggaaatg 120gtggtcttct tgagcgtatg anagacaccg gagttgatgt tatagggcta gactggacag 180tggatatggc agatggaaga agaagattgg gtagtgggat aggtgttcag ggaaatgtgg 240accctgccta cttattctcc cctcttgatg ccctgactga ag 282135256DNAGlycine max 135gggggatcct gttagtcgtc ctccggcatg gatgatgcgc caggccggaa ggtacatggc 60tgtttacaaa aagcttgctg agaaatatcc atccttccga gagaggtcag agacaactga 120tctcattgtg gaaatttctt tgcagccttg gaatgctttc aggcctgatg gagtaattat 180cttctcggac atccttacac cacttcctgc gtttggagtt gattttgaca tagaagaagt 240aaggggacct gttata 256136386DNAGlycine maxunsure at all n locations 136ttcaggctca gccgcatagt taaggaaccg aaactccaca taggaatcac ttggtttctt 60tgctctcccc caacccaatg gctacttcca ttaacagcag tgctctgggg tggaaacatt 120catccttctt cgtacaatcc aataatggct tcaacgttgc ttcgcctcct ttcaaaccaa 180agccgncacg ctcctccaac ttttctctct attgctctgc cgcctcctct tcttctgatc 240cactgttggt taaggctgct aggggagatc ctgttagtcg tcctccagca tggatgatgc 300gccaggcagg aaggtacatg gctgtttaca aaaatcttgc tgagaaatat ccatccttcc 360gagagaggtc agagacaact gaactc 386137291DNAGlycine max 137aggttttaca tccaattgac ctggacaggc ttaaatttgt tggagattca ctaaagatac 60tgcgccaaga ggttggtggt catgcagctg ttttgggttt tgtgggagca ccttggacaa 120tagcaacata tatagtggaa gggggtacaa cacgcacata tacaaccatt aagagcatgt 180gccacactgc cccacatgta ttgaggactt tgctttctca tttgacgcag gcaatagctg 240attacgttat tttccaagtg gagtctgggg ctcattgcat acaaatattt g 291138288DNAGlycine maxunsure at all n locations 138gcgccaagag gttggtggtc atgcagctgt tttgggtttt gtgggagcac cttgggacaa 60tagcaacata tatagtggaa gggggtacaa cacgcacata tacaaccatt aagagcatgt 120gccacactgc cccacatgta ttgaggactt tgctttctca tttgacgcag gcaatagctg 180attacgttat tttccaagtg gagtctgggg ctcattgcat acaaatattt gattcatgnc 240ngtggacaat accacctgaa atgtgggaac gctggtcaaa gccttata 288139261DNAGlycine max 139aaagatactg cgccaagagg ttggtggtca tgcagctgtc ttgggttttg tgggagcacc 60ttggacaata gcaacatata tagtggaagg gggtacaaca cgcacatata caaccattaa 120gagcatgtgc cacactgccc cacatgtatt gaggactttg ctttctcatt tgacgcaggc 180aatagctgat tacgttattt tccaagtgga gtctggggct cattgcatac aaatattaga 240tcatggggtg gacaactacc a 261140213DNAGlycine max 140gacaatagca acatatatag tggaaggggg tacaacacgc acatatacaa ccattaagag 60catgtgccac actgccccac atgtattgag gactttgctt tctcatttga cgcaggcaat 120agctgattac gttattttcc aagtggagtc tggggctcat tgcatacaaa tatttgattc 180atggggtgga caactaccac ctgaaatgtg gga 213141236DNAGlycine max 141tgttgaaaga cccccggttt ggctcatgag gcaagcaggg aggtacatga agagttacca 60aaccatctgt gagaaatatc cttcattccg tgaaagatct gaaaatgttg atctcgtggt 120ggaaatttct ctgcaaccat ggcatgtttt taagcccgat ggagtgattt tattctcaga 180cattcttacc ccactttctg gaatgaatat accctttgat attgtgaagg gtaagg 236142263DNAGlycine max 142tttggctcat gaggcaagca gggaggtaca tgaagagtta ccaaaccatc tgtgagaaat 60atccttcatt ccgtgaaaga tctgaaaatg ttgatctcgt ggtggaaatt tctctgcaac 120cgtggcatgt tttcaagcct gatggagtga ttttattctc agacattctt accccacttt 180ctggaatgaa tatacccttt gatattgtga agggtaaggg tcctgttata tttgatccta 240ttcacacatc tgcccaggtt gat 263143258DNAGlycine max 143gcttttgcta aatgcagttc gcgggataga tgttgaaaga cccccggttt ggctcatgag 60gcaagcaggg aggtacatga agagttacca aaccatctgt gagaaatatc cttcattccg 120tgaaagatct gaaaatgtga tctcgtggtg gaaatttctc tgcaaccgtg gcatgttttc 180aagcctgatg gagtgatttt attctcagac attcttaccc cactttctgg aatgaatata 240ccctttgata ttgtgaag 258144262DNAGlycine max 144caaacatgct ttgcgtcaac actgccttca cctctttctt gcccagaaaa tcaatttgct 60tcttttcctc caaatcaacc accccaattt cctgcaccct ccaaggaaca gttgcagaac 120caaaatctac agctgctggt gaacctcttt tgctaaatgc agttcgtggg atagatgttg 180aaagaccccc ggtttggctc atgaggcaag cagggaggta catgaagagt taccaaacca 240tctgtgagag atatccttca tt 262145283DNAGlycine max 145acttgttatc tatacagatg ttgcattaga tccttattca tcagatgggc atgatggcat 60agttagagaa gatggagtta ttatgaatga tgagacagtt catcagctat gtaaacaagc 120tgtagcccag gcccaagctg gagcagatgt tgtccagtct agtgatatga tggatggtcg

180ggtaggagca ctgcgtgcag ctctggatgc tgaaggcgtt cagcatgtat ctataatgtc 240ctatacagca aagtatgcaa gttcttttta tggtccattt aga 283146316DNAGlycine max 146ctgagatgcg ggaggatgaa tctgaaggag ctgacattct cttggtgaag cctggtcttc 60cttacttgga tatcataagg ctgctcaggg ataattctcc tttgccaatt gcagcatacc 120aggtttctgg tgaatatgca atgataaagg ctgccggtgc tctcaaaatg atagacgaag 180aaaaggttat gatggagtca ctgatgtgcc tccgaagggc cggtgctgat atcatcctca 240catattctgc tctgcaagct gccagatgtt tgtgtggaga gaagagtgaa gttctctgat 300tatgtagggc gttgtt 316147271DNAGlycine max 147tcgccggtaa ggttccgccg gcgcctcccg tgccgcccag accggcggct cccggttgga 60acaccggtgg ttccttcact tccacaccac cggcgtcctc gtcggaaccg gaagtcgccg 120gcgcttcggt cggcttttca ggaaacgagc atttcgccgg cgaatttcgt gtatccgctt 180ttcattcacg aaggtgaaga ggatactcca attggggcta tgcctggatg ctacaggctt 240gggtggaggc atggacttgt agaagaggtt g 271148275DNAGlycine maxunsure at all n locations 148aagcctggtc ttccttactt ggatatcata agtctgctca gggataattc tcctttgcca 60attgcagcat accaggttct tttctttgcc cattctagca ctaggcaaaa cgtttctgat 120aaaaagttga tcagatattc aatacatttt aaccagtgga attctgcntt aagcttgctg 180caagtgacag angtctatac gtagtagaca aatatcacac ctctagttta atatcaggct 240gaggtacaag tttatggttg ctttaacagt tattg 275149191DNAGlycine maxunsure at all n locations 149ccggtgctga tatcatcctc acatattctg ctctgcaagc tgccagatgt ttgtgtggag 60agaagaggtg aagttctctg attatgcagg gcgttgttca tgtagaaggt tgaagagttt 120anaaanccca gtnccggngn tncgggnnnt cnnaaaattt taaaagggnc cccgcggttt 180ntcnaaaang a 191150250DNAGlycine max 150aggagatgaa gcatacagtg aaaatggttt agtgcctcgg acaatacgtt tgctcaagga 60taagttacca gaccttggta accaatccag aggtggaata aaatcctaat ccgtcagatg 120ggcatgatgg catagtaaga gaagatgaag taataatgat tatgagacag gtcatcagcc 180atggtaacaa gctgtagacc aaggccaagc tggagcagat gttgtcagtc ctagtgatat 240gatggatggt 250151357DNAGlycine max 151acggctgcga caagacgaga taatgtggct gattggtaac gtagtgaatc ctgtgcatac 60atccgctcgt agcctcttcc tgcgactctc ttctcagtgg gtctccgtat tctccctcaa 120tcctattaac cttttcttct ttcatttccc accccattct ataatcaatc agtgtcaatg 180gcttcttcaa tcgctaatgc gccttctgcg ttcaattctc agtactactt tggtctcaga 240acgccactga ggtccttcaa cttttcttct cctcaagctg ccaaacttcc acgctcgcat 300tgccttttcg tcgtcagagc ctccgattcg gtcttcgaaa ccgccgttgt cgccggt 357152418DNAGlycine max 152agcccaggcg tcagtacggc tgcgagaaga cgacagaagg ggatggttga ctggttgttt 60tttaaattgc atgaaacatt tatttgttct tatagaaaaa gttacaagta agtcttcact 120gcaagtagaa gatattggat ccagttccag ggttgaactc catacgatta ttttttaata 180gaaaaattga ctgtgacgta gctgtggagg acacgattgg taaagtattg aatccttcct 240gcgactcttt tctcattggt tcactgtgtt ctccaaacac atctcagaat ctcttgtatt 300attattcaat caatcaatgg cttcttcaat ccctaatgga cctccctctg cgttgaattc 360ccagttctac gatgatctca gaccgccaca gaggaccttc aacttttcct ttcttcaa 418153243DNAGlycine max 153agcccaagcg tcagtacagc tgcgagagga ggacagaagg ggattctaca atcaatcaat 60ggcaatggct tcatcaatcc ctaatgcgcc ttctgcgttc aattctcaaa gctacgttgg 120tctcaggtcg ccactgagga ccttcaactt ttcttctcct caaggtggca aaaatcctcg 180ctcccaacgc cttttcgacg tcagagcctc cgaatccgag ttccaagccg ccgttgtccc 240cgg 243154277DNAGlycine maxunsure at all n locations 154cgcagtcnga ggancctcca cagatatnca nctcttaatg tgcaggaana tttccgnggc 60aatgtcnana caaggttaan aaagctcaat gagggggttg tccaagctac actattagca 120ttnnctggac tcaaacgctt aatatgacag anaatgtgac ttcaatccta tcantagatg 180atatgcttcc agctgttgnc caaggtgcca ttggaattgc ctgtagaagt gatgnnnata 240anatggcaga atacattgat tcacttaatc atganga 277155285DNAGlycine max 155tatgagatga agcatacagt gaaaatggtt tagtgcctcg gacaatacgt ttgctcaagg 60ataagtaccc agaccttgtt atctatacag atgttgcatt agatccttat tcgtcagatg 120ggcatgatgg catagttaga gaagatggag ttattatgaa tgatgagaca gttcatcagc 180tatgtaaaca agctgtagcc caggcccaag ctggagcaga tgttgtcagt cctagtgata 240tgatggatgg tcgggtagga gcactgcgtg cagctcttga tgctg 285156275DNAGlycine max 156acggctgcga gaagacgaca gaaggggatg ctttgaagtc tcccacagga gatgaagcat 60acaatgaaaa tggtttagtg cctcgaacaa tacgtttgct caaggataag tacccagacc 120ttgttatcta tacagatgtt gcattagatc cttattcatc agatgggcat gatggcatag 180ttagagaaga tggagttatt atgaatgatg agacagttca tcagctatgt aaacaagctg 240tagcccaggc ccaagctgga gcagatgttg tcagt 275157262DNAGlycine max 157ttttagtctc ccacaggaga tgaagcatac aatgaaaatg gtttagtgcc tcgaacaata 60cgtttactca aggataagta cccagacctt gttatctata cagatgttgc attagatcct 120tattcatcag atgggcatga tggcatagtt agagaagatg gagttattat gaatgatgag 180acagttcatc agctatgtaa acaagctgta gcccaggtca tatgactgtc ttctataaac 240attttcaact gtaggcagtt ac 262158289DNAGlycine max 158gaaaaggtta tgatggagtc actgatgtgc ctccgaaggc cggtgctgat atcatcctca 60catattctgc tctgcaagct gccagatgtt tgtgtggaga gaagaggtga agttctctga 120ttatgtaggg cgttgttcat gtagaaggtt gaagagttta taataccagt atctgctgga 180ttttggttat tgtaaattgt ttaagaggga catggaggtt tgtgtataga gagacattca 240taataaaata ttatggcctc gtttgattta atatatgtaa ggacataat 289159255DNAGlycine maxunsure at all n locations 159ggttatgatg gagtcactga tgtgcctccg aagggccggt gctgatatca tcctcacata 60ttctgctctg caagctgcca gatgtttgtg tggagagaag aggtgaagtt ctctgattat 120gtagggcgtt gttcatgtag aaggttgaag agtttataat accagtatct gctggatttt 180ggttattgta aattgtttaa gagggacatg gnggtttgtg tatagagaga cattcctaat 240taaatattag ggccc 255160262DNAGlycine maxunsure at all n locations 160tcgggtaggn gcactgcgtg cagctctgga tgctgaaggc tttcagcatg tttctataat 60gtcctataca gcaaagtatg caagttcttt tnatggtcca tttagagagg cactagactc 120aaacccccgg tttggagaca agaaaactta tcagatgaac ccagctaatt acagagaggc 180tctgactgag atgcgggagg atgaatctga aggagctgac attctcttgg tgaagcctgg 240tcttccttac ttggatatca ta 262161253DNAGlycine max 161gacagttcat cagctatgta aacaagctgt agcccaggcc caagctggag cagatgttgt 60cagtcctagt gatatgatgg atggtcgggt aggagcactg cgtgcagctc tggatgctga 120aggctttcag catgtttcta taatgtccta tacagcaaag tatgcaagtt ctttttatgg 180tccatttaga gaggcactag actcaaaccc ccggtttgga gacaagaaaa cttatcagat 240gaacccagct aat 253162249DNAGlycine max 162gttgtcagtc ctagtgatat gatggatggt cgggtaggag cactgcgtgc agctctggat 60gctgaaggct ttcagcatgt ttctataatg tcctatacag caaagtatgc aagttctttt 120tatggtccat ttagagaggc actagactca aacccccggt ttggagacaa gaaaacttat 180cagatgaacc cagctaatta cagagaggct ctgactgaga tgcgggagga tgaatctgaa 240ggagctgac 249163248DNAGlycine max 163gacagttcat cagctatgta aacaagctgt agcccaggcc caagctggag cagatgttgt 60cagtcctagt gatatgatgg atggtcgggt aggagcactg cgtgcagctc tggatgctga 120aggctttcag catgtttcta taatgtccta tacagcaaag tatgcaagtt ctttttatgg 180tccatttaga gaggcactag actcaaaccc ccggtttgga gacaagaaaa cttatcagat 240gaacccag 248164414DNAGlycine max 164acccacgcgt ccgtacggct ggagaagacg acagaagggg attctataat caatcaatgg 60caatggcttc ttcaatccct aatgcgcctt ctgcgttcaa ttctcagagc tacgttggtc 120tcagagcgcc actgaggacc ttcaactttt cttctcctca agctgccaaa attcctcgct 180cccaacgcct tttcgtcgtc agagcctccg attcggagtt cgaagccgcc gttgtcgccg 240gtaaggttcc gccggcgcct cccgtgccgc ccagaccggc ggctccggtt ggaacaccgg 300tggttccttc acttccactt caccggcgtc ctcgtcggaa ccggaagtcg ccggcgcttc 360ggtcggcttt tcaggaaacg agcatttcgc cggcgaattt cgtgtatccg cttt 414165394DNAGlycine max 165tacggctgcg agaagacgac agaaggggat aatcaatcaa tggcaatggc ttcttcaatc 60cctaatgcgc cttctgcgtt caattctcag agctacgttg gtctcagagc gccactgagg 120accttcaact tttcttctcc tcaagctgcc aaaattcctc gctcccaacg ccttttcgtc 180gtcagagcct ccgattcgga gttcgaagcc gccgttgtcg ccggtaaggt tccgccggcg 240cctcccgtgc cgcccagacc ggcggctccg gttggaacac cggtggttcc ttcacttcca 300cttcaccggc gtcctcgtcg gaaccggaag tcgccggcgc ttcggtcggc ttttcaggaa 360acgagcattt cgccggcgaa tttcgtgtat ccgc 394166283DNAGlycine maxunsure at all n locations 166gcttcttcaa tccctaatgc gccttctgcg ttcaattctc agagctacgt tggtctcaga 60gcgccactga ggaccttcaa cttttcttct cctcaagctg ccaaaattcc tcgctcccaa 120cgccttttcg tcgtcagagc ctccgattcg gagttcgnag ccgccgttgt cgccggtaag 180gttcncccgg cgcctcccgt gccgcccaga ccggcggctc cggttggaac accggtggtt 240ccttcacttc cacttcaccg gcgtcctcgt cggaaccgga agt 283167286DNAGlycine maxunsure at all n locations 167aatccctaat gcgccttctg cgttcaattc tcagagctac gttggtctca gagcgccact 60gaggaccttc aacttttctt ctcctcaagc tgccaaaatt cctcgctccc aacgcctttt 120cgtcgtcaga gcctccgatt cggagttcga agccgncgtt gtcgccggta aggttccgcc 180ggngcctccc gtnccgccca gaccggcggc tccggttgga acaccggtgg ttccttcact 240tccacttcac cggcgtcctc gtcggaaccg gaagtcgcgg cgcttt 286168278DNAGlycine max 168cttcaatccc taatgcgcct tctgcgttca attctcagag ctacgttggt ctcagagcgc 60cactgaggac cttcaacttt tcttctcctc aagctgccaa aattcctcgc tcccaacgcc 120ttttcgtcgt cagagcatcc gattcggagt tcgaagccgc cgttgtcgcc ggtaaggttc 180cgccggcgcc tcccgtgccg cccagaccgg cggctccggt tggaacaccg gtggttcctt 240cacttccact tcaccggcgt cctcgtcgga accggaag 278169268DNAGlycine max 169ggcttcttca atccctaatg cgccttctgc gttcaattct cagagctacg ttggtctcag 60agcgccactg aggaccttca acttttcttc tcctcaagct gccaaaattc ctcgctccca 120acgccttttc gtcgtcagag cctccgattc ggagttcgaa gccgccgttg tcgccggtaa 180ggttccgccg gcgcctcccg tgccgcccag accggcggct ccggttggaa caccggtggt 240tccttcactt ccacttcacc ggcgtcct 268170356DNAGlycine max 170attgaatcct gtgcatacat cctcacttat cctcttcctg cgactctctt ctcattggtt 60ctccgtattc tccctcaatc ctattaacct tttcttcttt catttcccac cccattctat 120aatcaatcaa tggcaatggc ttcttcaatc cctaatgcgc cttctgcgtt caattctcag 180agctacgttg gtctcagagc gccactgagg accttcaact tttcttctcc tcaagctgcc 240aaaattcctc gctcccaacg ccttttcgtc gtcagagcct ccgattcgga gttcgaagcc 300gccgttgtcg ccggtaaggt tccgccggcg cctcccgtgc cgcccagacc ggcggc 356171287DNAGlycine max 171gcttcttcaa tccctaatgc gccttctgct gttcaatgtc tcgagagctc acgttcgggt 60ctccagcagc gaccacttgc aggacgcttg cagacgtttt gcttagctcc tacgaagctt 120ggcgcaaata ttgcctgcgc tacccatacg ccttttacgt cgtcagagcc tccgattcgg 180agttcgaagc cgccgttgtc gccggtaagg ttccgccggc gcctcccgtg ccgcccagac 240cggcggctcc ggttggaaca ccggtggttc cttcacttcc acttcac 287172259DNAGlycine max 172atggcaatgg cttcttcaat ccctaatgcg ccttctgcgt tcaattctca gagctacgtt 60ggtctcagag cgccactgag gaccttcaac ttttcttctc ctcaagctgc caaaattcct 120cgctcccaac gccttttcgt cgtcagagcc tccgattcgg agttcgaagc cgccgttgtc 180gccggtaagg ttccgccggc gcctcccgtg ccgcccagac cggcggctcc ggttggaaca 240ccggtggttc cttcacttc 259173258DNAGlycine maxunsure at all n locations 173ggcttcttca atccctaatg cgccttctgc gttcaattct cagagctacg ttggtctcag 60agcgccactg aggaccttca acttttcttc tcctcaagct gccaaaattc ctcgctccca 120acgccttttc gtcgtcagag cctccgattc ggagttcgaa gccgccgttg tcgccggtaa 180ggttccgccg gcgcctcccg tgncgcccag accggcggct ccggttggaa caccggtggt 240tccttcattc cattcacc 258174234DNAGlycine max 174ggcttcttca atccctaatg cgccttctgc gttcaattct cagagctacg ttggtctcag 60agcgccactg aggaccttca acttttcttc tcctcaagct gccaaaattc ctcgctccca 120acgccttttc gtcgtcagag cctccgattc ggagttcgaa gccgccgttg tcgccggtaa 180ggttccgccg gcgcctcccg tgccgcccag accggcggct ccggttggaa cacc 234175251DNAGlycine maxunsure at all n locations 175gcttcttcaa tccctaatgc gccttctgcg ttcaattctc agagctacgt tggtctcaga 60gcgccactga ggaccttcaa cttttcttct cctcaagctg ccaaaattcc tcgctcccaa 120cgccttttcg tcgtcagagc ctccgattcg gagttcgang ccgccgttgt cgccggtnag 180gttccgccgg cgcntcccgt nccgcccaga ccggcggctc cggttggaac aaccggtggt 240tccttcactt c 251176279DNAGlycine max 176atccctaatg cgccttctgc gttcaattct cagagctacg ttggtctcag agcgccactg 60aggaccttca acttttcttc tcctcaagct gccaaaattc ctcgctccca acgccttttc 120gtcgtcagag cctccgattc ggagttcgaa gccgccgttg tcgccggtaa ggttccgccg 180gcgcctcccg tgccgcccag accggcggct ccggttggaa caccggtggt tccttcactt 240ccacttcacc ggcgtcctcg tcggaaccgg aagtcgccg 279177266DNAGlycine max 177ggcttcttca atccctaatg cgccttctgc gttcaattct cagagctacg ttggtctcag 60agcgccactg aggaccttca acttttcttc tcctcaagct gccaaaattc ctcgctccca 120acgccttttc gtcgtcagag cctccgattc ggagttcgaa gccgccgttg tcgccggtaa 180ggttccgccg gcgcctcccg tgccgcccag accggcggct ccggttggaa caccggtggt 240tccttcactt ccacttcacc ggcgtc 266178287DNAGlycine max 178atcctattaa ccttttcttc tttcatttcc caccccattc tatagtcaat caatggcaat 60ggcttcttca atccctaatg cgccttctgc gctcaattct cagagctacg ttggtctcag 120agcgccactg aggaccttca acttttcttc tcctcaagct gccaaaattc ctcgctccca 180acgccttttc gtcgtcagag cctccgattc ggagttcgaa gccgccgttg tcgccggtaa 240ggttccgccg gcgcctcccg tgccgcccag accggcggct ccggttg 287179236DNAGlycine max 179caatggcaat ggcttcttca atccctaatg cgccttctgc gttcaattct cagagctacg 60ttggtctcag agcgccactg aggaccttca acttttcttc tcctcaagct gccaaaattc 120ctcgctccca acgccttttc gtcgtcagag cctccgattc ggagttcgaa gccgccgttg 180tcgccggtac agttccgccg gcgctcccgt gccgcccaga ccggcggctc cggttg 236180395DNAGlycine maxunsure at all n locations 180tacggatgcg agaagacgac agaaggggga ttggtaaagt attgaatcct gtgcatacat 60cctcacttat cctcttcctg cgactctctt ctcattggtt ctccgtattc tccctcaatc 120ctattaacct tttcttcttt catttcccac cccattctat aatcaatcaa tggcaatggc 180ttcttcaatc cctaatgcgc cttctgcgtt caattctcag agctacgttg gtctcagagc 240gccactgagg accttcaact tttcttctcc tcaagctgcc aaaattcctc gctcncaacg 300ccttttcgtc gtcagagcct ccgattcgga gttcgaagcc gccgttgtcg ccggtaaggt 360tccgccggcg cctcccgtgc cgcccagacc ggcgg 395181227DNAGlycine max 181tggcttcttc aatccctaat gcgccttctg cgttcaattc tcagagctac gttggtctca 60gagcgccact gaggaccttc aacttttctt ctcctcaagc tgccaaaatt cctcgctccc 120aacgcctttt cgtctcagag cctccgattc ggagttcgaa gccgccgttg tcgccggtaa 180ggttccgccg gcgcctcccg tgccgcccag accggcggct ccggttg 227182271DNAGlycine maxunsure at all n locations 182ggcttcttca atccctaatg cgccttctgc gttcaattct cagagctacg ttggtctcag 60agcgccactg aggaccttca acttttcttc tcctcaagct gccaaaattc ctcgctccca 120acgccttttc gtcgtcagag cctccgattc ggagttcgaa gcagccgttg tcgccggtaa 180ggttccgccg gngcttccnt gccgnacaga ccggcgggtc cngttggnac aacggtggtt 240ccttaattcc actnancggc gtcctntcng a 271183256DNAGlycine max 183cggctcgaga aaattgactg tcacgtagct gaagctgatt gagctacgtt ggtctcagag 60cgccactgag gaccttcaac ttttcttctc ctcaagctgc caaaattcct cgctcccaac 120gccttttcga cgtcagagcc tccgattcgg agttcgaagc cgccgttgtc gccggtaagg 180ttccgccggc gcctcccgtg ccgcccagac cggcggctcc ggttggaaca ccggtggttc 240cttcacttcc acttca 256184246DNAGlycine max 184accttgtctt ctttcatttc ccaccccatt ctataatcaa tcaatggcaa ttgcttcttc 60aatccctaat gcgccttctg cgttcaattc tcagagctac gttggtctca gagcgccact 120gaggaccttc aactttgctt ctcctcaagc tgccaaaatt cctcgctccc aacgcctttt 180cgtcgtcaga gcctccgatt cggagttcga agccgccgtt gtcgccggta agttccgccg 240gcgctt 246185253DNAGlycine max 185cgactctctt ctcattggtt ctccgtattc tccctcaatc ctattaacct tttcttcttt 60catttcccac cccattctat aatcaatcaa tggcaatggc ttcttcaatc cctaatgcgc 120cttctgcgtt caattctcag agctacgttg gtctcagagc gccactgagg accttcaact 180tttcttctcc tcaagctgcc aaaattcctc gctcccaacg ccttttcgtc gtcagagcct 240ccgattcgga gtt 253186148DNAGlycine max 186ctgcgttcaa ttctcagagc tacgttggtc tcagagcgcc actgaggacc ttcaactttt 60cttctcctca agctgccaaa attcctcgct cccaacgcct tttcgtcgtc agagcctccg 120attcggagtt cgaagccgcc gttgtcgc 148187271DNAGlycine max 187cggctcgagg ctgaagctga ttggtaaagt attgaatcct gtgcatacat cctcacttat 60cctcttcctg cgactctctt ctcattggtt ctccgtattc tccctcaatc ctattaacct 120tttcttcttt catttcccac ccattctata atcaatcaat ggcaatggct tcttcaatcc 180ctaatgcgcc ttctgcgttc aattctcaga gctacgttgg tctcagagcg ccactgagga 240ccttcaactt ttcttctcct caagctgcca a 271188104DNAGlycine max 188atggcttctt caatccctaa tgcgccttct gcgttcaatt ctcagagcta cgttggtctc 60agagcgccac tgaggacctt caacttttct tctcctcaag ctgc 10418964DNAGlycine max 189agcttcttca atccctaatg cgccttctgc gttcaattct cagagctacg ttggtctcag 60agcg 64190266DNAGlycine max 190tcggctcact cgagcgaatc ggctcaggaa aattgactgt gacgtagcac atcctgattg 60gtaaactatt gaatcctgtg catacatcct cacttatcct cttcctgcga ctctcttctc 120cttggttctc cgtattctcc ctcaatccta ttaacctttt cttctttcat ttcccacccc 180attctataat caatcaatgg caatggcttc ttcaatccct aatgcgcctt ctgcgttcaa 240ttctcagagc tacgttggtc tcagag 266191264DNAGlycine max 191ctcatataga aaattgactg tgacgttgct gaagctgatt ggtaaagtat tgaatcctgt 60gcatacatcc tcacttatcc tcttcctgcg actctcttct cattggttct ccgtattctc 120cctcaatcct attgaccttt tcttctttca tttcccaccc cattctataa tcaatcaatg 180gcaatggctt cttcaatccc taatgcgcct tctgcgttca attctcagag ctacgttggt 240ctcagagcgc cactgaggac cttc

264192335DNAGlycine maxunsure at all n locations 192atatgctnnc cagctgttgc ccaaggtgcn attggaatag cctgtagaag taacgatgat 60aaaatgnnca gaatacctcn ncttcattga atcatgaaga aacaagacta gcagtttgct 120gtgaaagagc cttccttgan aagtagaagg atntgccgna nnctattgca ggctatgcta 180gcagaaacga ggatggcaat tgcttgttta gaggatagtt gcttcccctg atggaacccg 240cgtgctcgaa actccagaat ggttcanatg ctttcgaaga tatgataaag atgggtaaga 300tgctggagag gagctctttc tcgagctgac ntgct 335193257DNAGlycine max 193gaacagcgaa atcgacatcg ctgtccattc gatgaaggat gttcctactt acttgcctga 60taaaacaatt ctgccatgta accttccgcg agaggatgtc agagatgcat ttatatcctt 120gactgcagct tccttagctg atcttccccc tgcaagtgtt attggtactg cttcgttaag 180gcgaaagtca cagatcctcc acagatatcc atctcttaat gtgcaggaaa atttccgtgg 240caatgtccaa acaaggt 257194269DNAGlycine maxunsure at all n locations 194cgtttaaata tgacggaaaa tgtgacttcg atcctatcaa ttgatgacat gcttccagct 60gttgcccaag gtgcaattgg aatagcctgt agaagtaatg atgataaaat ggcggaatac 120cttgcttcac tgaatcatga agaaacaaga ctagcagttt cctgcgaaag angcttcctt 180gaaaagttgg aagggtctgc cgcactccta ttgcaggcta tgctagcaga aatgaggatg 240gcaattgctt gtttagagga ttagttgca 269195259DNAGlycine max 195tgatgataaa atggcggaat accttgcttc actgaatcat gaagaaacaa gactagcagt 60ttcctgtgaa agatccttcc ttgaaaagtt ggaagggtct tgccgcactc ctattgcagg 120ctatgctagc agaaatgagg atggcaattg cttgtttaga ggattagttg catcccctga 180tggaatccgt gtgcttgaaa cttccagaat tggcccatat gcgttcgcag atatgataaa 240gatgggtaag gatgctgga 259196205DNAGlycine max 196cttaagtatg acagaaaatg tgacttcaat cctatcaatt gatgatatgc ttccagctgt 60tgcccaaggt gctattggaa tagcatgtag aagtgatgac gataaaatgg cggaatacat 120tgctacactt aatcatgaag aaacaagact agcagttgtc tgtgagaggg cctttcttca 180gactttggat gggtctgccg cactc 205197271DNAGlycine max 197ctgcttcgtt aaggcgaaag tcacagatcc tccacagata tccatctctt aatgtgcagg 60aaaatttccg tggcaatgtc caaacaaggt taagaaaact caatgagggg gttgtccaag 120ctacactatt agcattagct ggactcaaac gcttaagtat gacagaaaat gtgacttcaa 180tcctatcaat agatgatatg cttccagctg ttgcccaagg tgccattgga attgcctgta 240gaagtgatga cgataaaatg gcagaataca t 271198287DNAGlycine max 198attggaattg cctgtagaag tgatgacgat aaaatggcag aatacattga ttcacttaat 60catgaagaaa caaggctagc agttgtctgt gaaagggcct ttcttcagac tttggatggg 120tcttgccgca ctcctattgc agggtatgct tgtagaaacg aggatggcaa ttgtttgttt 180agaggattag ttgcttcccc tgatggaacc agagtgctag agacatccag ggttggtcca 240tatgctgttg aagatatgat tgagatgggt aaggatgctg gcaagga 287199276DNAGlycine max 199attgggaatt gcctgtagaa gtgatgacga taaaatggca gaatacattg attcacttaa 60tcatgaagaa acaaggctag cagttgtctg tgaaagggcc tttcttcaga ctttggatgg 120gtcttgccgc actcctattg cagggtatgc ttgtagaaac gaggatggca attgtttgtt 180tagaggatta gttgcttccc ctgatggaac cagagtgcta gagacatcca gggttggtcc 240atatgctgtt gaagatatga ttgagatggg taagga 276200285DNAGlycine max 200attggaattg cctgtagaag tgatgacgat aaaatggcag aatacattga ttcacttaat 60ccatgaagaa acaaggctag cagttgtctg tgaaagggcc tttcttcaga ctttggatgg 120gtcttgccgc actcctattg cagggtatgc ttgtagaaac gaggatggca attgtttgtt 180tagaggatta gttgcttccc ctgatggaac cagagtgcta gagacatcca gggttggtcc 240atatgctgtt gaagatatga ttgagatggg taaggatgct ggcaa 285201259DNAGlycine max 201gtgaaagggc ctttcttcag actttggatg ggtcttgccg cactcctatt gcagggtatg 60cttgtagaaa cgaagatggc aattgtttgt ttagaggatt agttgcttcc cctgatggaa 120ccagagtgct agagacatcc agggttggtc catatgctgt tgaagatatg attgagatgg 180gtaaggatgc tggcaaggag cttctgtctc gggctggacc taacttcttc agtagttagc 240agcagatgat taaagtgtg 259202285DNAGlycine maxunsure at all n locations 202gcagacagaa gcgaacgnaa cggggttgcc tcaacaattc gctgttgttg ttctcttctc 60ttctctttga catgaatact ctttcttcca cgctccatgg cggcaggctt ccccgctcag 120cttcgaaaac caaaaccgca tctctctcca aatgccatcg catttgggtc accaaagctt 180ctgttgccgt tgagcaacaa actaaggtcg ctctcatcag aattggtacc agaggaagtc 240cactagctct agcacaagca tatgagacca gagacaaact catgg 285203282DNAGlycine max 203agcagacaga agcgagcgaa acggggttgc ctcaacaatt cgctgttgtt gttctcttct 60cttctctttg acatgaatac tctttcttcc acgctccatg gcgggaggct tccccgctca 120gcttcgaaaa ccaaaaccgc atctctctcc aaatgccatc gcatttgggt caccaaagct 180tctgttgccg ttgagcaaca aactaaggtc gctctcatca gaattggtac cagaggaagt 240ccactagctc tagcacaagc atatgagacc agagacaaac tc 282204251DNAGlycine max 204ccgaacgaaa cggggttgcc tcaacaattc gctgttgttg ttctcttctc ttctctttga 60catgaatact ctttcttcca cgctccatgg cgggtggctt ccccgctcag cttcgaaaac 120cacaaccgca tctctctcca aatgccatcg catttgggtc accaaagctt ctgttgccgt 180tgagcaacaa actaaggtcg ctctcatcag aattggtacc agaggaagtc cactagctct 240agcacaagca t 251205327DNAGlycine max 205atcggcaagg taaggcaatt gaagttgtga aatggagact gtctgctctg cattggtgtt 60cccatctttc agaatcacaa cttcagcttt ctccaaatgt ggcatcaggg cttccattgc 120cgttgagcaa caaacttcgc agactaaggt tgctctcctc aaaattggta ccagaggaag 180tccactagct ctggctcagg catatgagac cagagacaag ctcatggcat cacatccaga 240gctagcggaa gaaggggcta ttcagattgt gataatgaaa acaactggtg acaaaatact 300atcacagcca cttgcagaca tcggcgg 327206390DNAGlycine max 206gaaatggaga ctctctgctc tgcattggtg ttcccatctt tcagaatcac aacttcagct 60ttctccaaat gtggcatcag ggctttcatt gccgttgagc aacatacttc gcagactaag 120gttgctctcc tcaaaattgg taccagagga agtccactag ctctggctca tgcatatgag 180accagagaca atctcatggc atcacatcca gagctagcgg atgaaggggc tattcagatc 240gtgataataa aaacaactgg tgacattata ctatcacagc cacttgcaga catcggcggt 300aagggcctgt ccacaatcga tatagacgag gcactcatta acggtgacat tgacatcgcc 360gttcactcta tgaaagatgt acccacttac 390207256DNAGlycine max 207cgttgctctc ctcagaattg gtaccagagg aagtccacta gctctggctc acgcatatga 60gaccagagac aagctcatgg catcacatgc agagctagca caagaagggg ctattcagat 120tgtaataatc aaaacaactg gtgacaaaat actatcacag ccacttgcag acattggtgg 180gaagggccta ttcacaaaag aaatagatga ggcactcata aacggtgaca ttgacatcgc 240tgtccactca atgaaa 256208289DNAGlycine maxunsure at all n locations 208ggagaccctc tgnctctgca ttggtgttcc catctttcag aatcagnact tcagctttct 60ccaaatgtgg catcagggcn tccattgccg ttgagcaaca aanttcccag actaaggttg 120ctctcctcag aattggtacc agaggaagtc cactagctct ggctcaggca tatgagacca 180gagacaagct catggcatca catgcagagc tagcagaaga aggggctatt cagnttgtaa 240taataanaac nactggtgac aanatactat cacagccact tgcagacat 289209259DNAGlycine maxunsure at all n locations 209agggcttcca ttgccgttga gcaacaaact tcccagacta aggttgctct cctcagaatt 60ggtaccagag gaagtccact agctctggct cncgcatatg agaccagaga caagctcatg 120gcatnccatg cagagctagc agaagaaggg gctattcaga ttgtaataat aaaaacaact 180ggtgacaaaa tactatcaca gccacttgca gacattggtg ggaagggcct attcacaaaa 240gaatagatga ggcatcata 259210268DNAGlycine max 210ctctctgctc tgcattggtg ttcccatatt tcagaatcac aacttcagct ttctccaaat 60gtggcatcag ggcttccatt gccgttgagc aacaaacttc gcagactaag gttgctctcc 120tcaaaattgg taccagagga agtccactag ctctggctca ggcatatgag accagagaca 180agctcatggc atcacatcca gagctagcgg aagaaggggc tattcagatt gtgataataa 240aaacaactgg tgacaaaata ctatcaca 268211270DNAGlycine max 211ggagactctc tgctctgcat tggtgttccc atctttcaga atcacaactt cagctttctc 60caaatgtggc atcagggctt ccattgccgt tgagcaacaa acttcgcaga ctaaggttgc 120tctcctcaaa attggtacca gaggaagtcc actagctctg gctcaggcat atgagaccag 180agacaagctc atggcatcac atccagagct agcggaagaa ggggctattc agattgtgat 240aataaaaaca actggtgaca aaatactatc 270212295DNAGlycine maxunsure at all n locations 212tggagaccct ctgctctgca ttggtgttcc catctttcag aatcagaact tcagctttct 60ccaaatgtgg catcagggct tccattgccg ttgagcaaca aacttcccag actaaggttg 120ctctcctcag aattggtacc agaggaagtc cactagctct ggctcaggca tatgagacca 180gagacaagct catggcatca catgcagagc tagcagaaga aggggctatt cagattgtat 240aataanaaca actggtgaca aaatatatca cagccattgc agacattggt gggag 295213267DNAGlycine max 213ctctctgctc tgcattggtg ttcccatctt tcagaatcac aacttcagct ttctccaaat 60gtggcatcag ggcttccatt gccgttgagc aacaaacttc gcagactaag gttgctctcc 120tcaaaattgg taccagagga agtccatagc tctggctcag gcatatgaga ccagagacaa 180gctcatggca tcacatccag agctagcgga agaaggggct attcagattg tgataataaa 240aacaactggt gacaaatact atcacag 267214251DNAGlycine max 214tggagactct ctgctctgca ttggtgttcc catctttcag aatcacaact tcagctttct 60ccaaatgtgg catcagggct tccattgccg ttgagcaaca aacttcgcag actaaggttg 120ctctcctcaa aattggtacc agaggaagtc cactagctct ggctcaggca tatgagacca 180gagacaagct catggcatca catccagagc tagcggaaga aggggctatt cagattgtga 240taataaaaac a 251215159DNAGlycine maxunsure at all n locations 215ccacttcagc tttctccaaa tgtggcatca gggcttccat tgccgttgag caacaaactt 60cccagactaa ggttgctctc ctcagaattg gtaccagagg aagtccacta gctctggctc 120aggcatatgn gaccagagac aagntcatgg catcacang 159216270DNAGlycine max 216gttcccatct ttcagaatca gaacttcagc tttctccaaa tgtggcatca gggcttccat 60tgccgttgag caacaaactt cccagactaa ggttgctctc ctcagaattg gtaccagagg 120aaggtaccct acccttaaaa ataacacctt tagcttctta tgagcatttc ttttaaagaa 180caagtctgtg aaaatattga gtcctgaatc tcttcaaaac tttgccctca ttttcaaatt 240tagttttcaa tgctagtttt atgacagaaa 270217147DNAGlycine max 217gtgaaatgga gaccctctgc tctgcattgg tgttcccatc tttcagaatc agaacttcag 60ctttctccaa atgtggcatc agggcttcca ttgccgttga gcaacaaact tcccagacta 120aggttgctct cctcagaatt ggtacca 147218253DNAGlycine maxunsure at all n locations 218ccaagaccga caacaaactc actcttacca agtccgagga agctttcgct gctgccaagg 60agcngatgcc tggaggtgtc aactccccag ttngtgcctt caaatccgtg ggtggtcaac 120caattgtgat tgattcagtc aaagggtctc gtatgtggga catcgacggc aatgagtaca 180ttgactacgt cggttcttgg ggtcccgcaa tcattggtca cgctgatgat caagtgcttt 240cagctctggt tgt 253219264DNAGlycine max 219tgcgtgcgtg agcgtcttac ctttccatta tcaaaatgac tgtttcagct atcacaggct 60cgcagtctca cctcttgcca tggttagcga tacctctttc ctctcccacg cgctctcgaa 120tcgtcgcaat ggccgtatcc gtcgtcccca agaccgacaa caaactcact cttaccaagt 180ccgaagcagc tttcgctgct gccaaggagc tgctgcctgg cggtgtcaac tccccagttc 240gtaccttcaa atccgtaggt ggtc 264220157DNAGlycine max 220ctcgtctgag ggctgttacc atggccatgc tgatcctttt cgtgttaagg caggtagtgg 60agttgccacc ttgggacttc ctgattctcc cggtgtcccc aaagctgaca ctgtggaaac 120ccttacagcg ccctacaatg atactgccgc cgtcgag 157221266DNAGlycine max 221aaacccgatt ttcataattt cttgcgcaag atcaccaagg agaacaatac ccttcttgtg 60tttgatgaag ttatgactgg gtttcgtttg tcatacggag gtgctcaaga gtattttggc 120ataactcctg atatacaact ctaggaaaga tcattggtgg aggtctgccg gtgggggctt 180atggagggag gagggatatt atggagaagg tggcaccagc tggcccaatg tatcaggctg 240ggaccttgag tgggaacctt tggcca 266222250DNAGlycine max 222aaaggagaaa ttgccgcagt tttcctcgaa cctgttgttg gaaacgctgg tttcattgtt 60cctaagcctg attttcatag tttcttgcgc aagatcacca aggagaacaa tacccttctt 120gtgtttgatg aagtcatgac tggatttcgt ttgtcatatg gaggtgctca agagtattat 180ggcataactc cagatataac aactctagga aagatcattg gtggaggtct gccggtaggg 240cttatggagg 250223256DNAGlycine max 223gctcaagagt attttggcat aactcctgat ataacaactc taggaaagat cattggtgga 60ggtctgccgg tgggggctta tggagggagg agggatatta tggagaaggt ggcaccagct 120ggcccaatgt atcaggctgg gaccttgagt gggaaccctt tggccatgac tgcaggaata 180cagaccctgc agcgtattaa ggagccagga acttatgagt acttggacaa aatcaccggt 240gagcttgttc agggca 256224288DNAGlycine maxunsure at all n locations 224tttaggnagc tgatgcctgg anggcgtgaa ctccccagtt cgtgncttca aatccgtggg 60tggtcaacca attgtgattg attcagtcaa agggtctcgt atgtgggata tcgatggcaa 120tgagtacatt gactacgttg gttcctgggg tcctgcaatc attggtcacg ctgatgatca 180ggtgcttgca gctctgggtg aaaccatgaa ganaggaacc agctttgggt gcaccctgtc 240tgctggaaaa cacttttggc agagctgggt tatcgatgcc gtncccca 288225283DNAGlycine maxunsure at all n locations 225attttgcaga tgccaaaaag agtgatacgg ccaagtttgc taggcccttt tggggaatgc 60tggcggaagg tgtctatttg gcaccttccc agnttgangc nggcttcacc agcttggcac 120atacttctgn tgacataaaa aagacgatan ccgctgntga gaaggttttc anggagntct 180gatggttaaa ttttgntttg ttgcaaattt aattntcgga gggtgaattt ttaggtcaat 240ttngattatt gttatggcag ttgctttcgn tatgatctgt atc 283226249DNAGlycine max 226gggtcctgca atcattggtc acgctgatga tcaggtgctt gcagctctgg gtgaaaccat 60gaagaaagga accagctttg gtgcaccctg tctgctggaa aacactttgg cagagctggt 120tatcgatgcc gtccccagca ttgaaatggt tcggtttgtc aattcaggca ctgaagcttg 180catgggtgcg ctccgtctgg cccgtgctta taccggaaga gagaagatca tcaagtttga 240gggctgtta 249227442DNAGlycine max 227ataaggcttt gcatttcatt tgagagagag agcgtcttac ctttccatta tcaaaatggg 60tgggtcggct atcacaggag cgaggctaac cctagggata gggttggcga tacctctttc 120ctctcccacg cgctctcgaa ccgtcgcaat ggccgtatcc gtcgacccca agaccgacaa 180caaactcact cttaccaagt ccgaggaagc tttcgctgct gccaaggtac gcatgacctc 240cctcttcctt ccttccttcc tcctttcaat tttgattttt gatttttgat ttcaggagct 300gatgcctgga ggtgtcaact ccccagttcg tgccttcaaa tccgtgggtg gtcaaccaat 360tgtgattgat tcagtcaaag ggtctcgtat gtgggacatc gacggcaatg agtacattga 420ctacgtcggt tcttggggtc cc 442228275DNAGlycine maxunsure at all n locations 228tcaaaatggc tgtttcggct atcacaggag cgaggctaac cctagggata gggttggcga 60tacctctttc ctctcccacg cgctctcgaa ccntcgcaat ggccgtatcc gtcgacccca 120agaccgacaa caaactcact cttaccaagt ccgaggaagc tttcgctgct gccaaggagc 180tgatgcctgg aggtgtcaac tccccagttc gtgccttcaa atccgtgggt ggtcaaccaa 240ttgtgattga ttcagtcaaa gggtctcgta tgtgg 275229261DNAGlycine max 229acccacgcgt ccgacggctg caagaggacg acagaagggg aaggctttgc atttcatttg 60agagagagag cgtcttacct ttccattatc aaaatggctg tttccgctat cacaggagcc 120aagctaaccc taaggataag gttggcgata cctccttcct ctcccaagcg ctctcgaacc 180gtcgcaatgg ccgtatccgt cgaccccaag accgacaaca aactcaatcc taccaagtcc 240gaagaagctt tcgctgctgc c 261230289DNAGlycine maxunsure at all n locations 230ggagaggata aggctttgca tttcatttga gaganagagc gtcttacctt tccattatca 60aaatggctgt ttcggctatc acaggagcga ggctaaccct agggataggg ttggcgatac 120ctctttcctc tcccacgcgc tctcgaaccg tcgcaatggc cgtatccgtc gaccccaaga 180ccgacaacaa actcactctt accaagtccg aggaagcttt cgctgctgcc aaggagctga 240tgcctggagg tgtcaactcc ccagttcgtg ccttcaaatc cgtgggtgg 289231252DNAGlycine max 231agcgtcttac ctttccatta tcaaaatggc tgtttcggct atcacaggag cgaggctaac 60cctagggata gggttggcga tacctctttc ctctcccacg cgctctcgaa ccgtcgcaat 120ggccgtatcc gtcgacccca agaccgacaa caaactcact cttaccaagt ccgaggaagc 180tttcgctgct gccaaggagc tgatgcctgg aggtgtcaac tccccagttc gtgccttcaa 240atccgtgggt gg 252232281DNAGlycine max 232ggctttgcat ttcatttgag agagagagcg tcttaccttt ccattatcaa aatggctgtt 60tcggctatca caggagcgag gctaacccta gggatagggt tggcgatacc tctttcctct 120cccacgcgct ctcgaaccgt cgcaatggcc gtatccgtcg accccaagac cgacaacaaa 180ctcactctta ccaagtccga ggaagctttc gctgctgcca aggagctgat gcctggaggt 240gtcaactccc cagttcgtgc cttcaaatcc gtgggtggtc a 281233276DNAGlycine max 233taaggctttg catttcattt gagagagaga gcgtcttacc tttccattat caaaatggct 60gtttcggcta tcacaggagc gaggctaacc ctagggatag ggttggcgat acctctttcc 120tctcccacgc gctctcgaac cgtcgcaatg gccgtatccg tcgaccccaa gaccgacaac 180aaactcactc ttaccaagtc cgaggaagct ttcgctgctg ccaaggagct gatgcctgga 240ggtgtcaact ccccagttcg tgccttcaaa tccgtg 276234276DNAGlycine max 234ttgcatttca tttgagagag agagcgtctt acctttccat tatcaaaatg gctgtttcgg 60ctatcacagg agcgaggcta accctaggga tagggttggc gatacctctt tcctctccca 120cgcgctctcg aaccgtcgca atggccgtat ccgtcgaccc caagaccgac aacaaactca 180ctcttaccaa gtccgaggaa gctttcgctg ctgccaagga gctgatgcct ggaggccgtc 240aatccccagt tcgtgccttc aaatccgtgg gtggtc 276235251DNAGlycine max 235tttgcatttc atttgagaga gagagcgtct tacctttcca ttatcaaaat ggctgtttcg 60gctatcacag gagcgaggct aaccctaggg atagggttgg cgatacctct ttcctctccc 120acgcgctctc gaaccgtcgc aatggccgta tccgtcgacc ccaagaccga caacaaactc 180actcttacca agtccgagga agctttcgct gctgcaagga gctgatgcct ggaggtgtca 240actccccagt t 251236271DNAGlycine max 236cggctcgaca aggctttgca tttcatttga gagagagagc gtcttacctt tccattatca 60aaatggctgt ttcggctatc acaggagcga ggctaaccct agggataggg ttggcgatac 120ctctttcctc tcccacgcgc tctcgaaccg tcgcaatggc cgtatccgtc gaccccaaga 180ccgacaacaa actcactctt accaagtccg aggaagcttt cgctgctgcc aaggagctga 240tgcctggagg tgtcaactcc ccagttcgtg c 271237257DNAGlycine max 237ggagaggata aggctttgca tttcatttga gagagagagc gtcttaactt tacattatca 60aaatggctgt ttcggctatc acaggagcga ggctaaatct agggataggg ttggcgatac 120ctctttcctc tcccacgcgc tctcgaaccg tcgcaatggc cgtatccgtc gaccccaaga 180ccgacaacaa actcactctt accaagtccg aggaagcttt cgctgctgcc aaggagctga 240tgcctggagg tgtcaac 257238153DNAGlycine maxunsure at all n locations 238acaggagcga ggctaaccct

agggataggg ttggcgatan ctctttcctc tcncactccg 60ctctcgaacc ntcgcaatgg ccgtatccgt cgaccccaag acngacaaca aactcactct 120taccaagtcc gaggaagctt tcgctgctgc caa 153239104DNAGlycine maxunsure at all n locations 239acggctgcga gaagacgaca gaagggggag cgtcttacct ttccattatc aaaatggcta 60tttcggctat cacaggagcg aggctaancc tagggatagg gttg 104240268DNAGlycine max 240ggctgggacc ttgagtggga accctttggc catgactgca ggaatacaga ccctgcagcg 60tattaaggag ccaggaactt atgagtactt ggacaaaatc accggtgagc ttgttcaggg 120cattattgaa gctgggaaga gggcaggcca tgcaatatgt ggtggtcata taagggggat 180gtttgggttt ttcttcacag aaggaccagt gtataatttt gcagatgcca aaaagagtga 240tacggacaag tttctaggtt cttttggg 268241256DNAGlycine max 241gaaggtggca ccagctggcc caatgtatca ggctgggacc ttgagtggga accctttggc 60catgactgca ggaatacaga ccctgcagcg tattaaggag ccaggaactt atgagtactt 120ggacaaaatc accggtgagc ttgttcaggg cattattgaa gctgggaaga gggcaggcca 180tgcaatatgt ggtggtcata taagggggat gtttgggttt ttcttcacag aaggaccagt 240gtataatttt gcagat 256242253DNAGlycine max 242ggcaccagct ggcccaatgt atcaggctgg gaccttgagt gggaaccctt tggccatgac 60tgcaggaata cagaccctgc agcgtattaa ggagccagga acttatgagt acttggacaa 120aatcaccggt gagcttgttc agggcattat tgaagctggg aagagggcag gccatgcaat 180atgtggtggt catataaggg ggatgtttgg gtttttcttc acagaaggac cagtgtataa 240ttttgcagat gcc 253243269DNAGlycine max 243ctcgagccgc tcgagccggt ctgctggaaa acactttggc agagctggtt atcaatgcgg 60tccccagcat tgcaatggtt cgctttgtca attcaggcac cgaagcttgc atgggtgcac 120tacgtctcgc ccgagcttat accggaagag agaagatcat caagtttgag ggctgttacc 180atggccatgc tgatcctttt cttgttaagg caggtagtgg agttgccacc ttgggacttc 240ctgattctcc cggtgtcccc aaagctgcc 269244266DNAGlycine max 244ctcgagccgc tcgagccggt ctgctggaaa acactttggc agagctggtt atcaatgcgg 60tacccagcat taccaatggt tcgctttgtc aattcaggca ccgaagcttg catgggtgca 120ctacgtctcg cccgagctta taccggaaga gagaagatca tcaagtttga gggctgttac 180catggccatg ctgatccttt tcttgttaag gcaggtagtg gagttgccac cttgggactt 240cctgattctc ccggtgtccc caaagc 266245266DNAGlycine max 245tcaagtttga gggctgttac cgtggccatg ctgatccttt tcttgttaag gcaggtagtg 60gagttgccac cttaggactt cctgattctc ccggtgtccc caaagctgcc acttttgaaa 120cccttacagc cccctacaat gacaccgagg ccattgagaa actcttcgag gccaacaaag 180gagaaattgc cgcagttttc ctcgaacctg ttgttggaaa cgctggtttc attgttccta 240agcctgattt tcatagtttc ttgcgc 266246238DNAGlycine max 246gttaccatgg ccatgctgat ccttttcttg ttaaggcagg tagtggagtt gccaccttgg 60gacttcctga ttctcccggt gtccccaaag ctgccacttt tgaaaccctt acagccccct 120acaatgacac tgccgccgtt gagaagctct ttgaggctaa caaaggagaa atcgctgctg 180ttttcctcga acctgttgtt ggaaacgctg gtttcattgt tcctaaaccg attttcat 238247232DNAGlycine max 247gggagatctg attgttaaat tttgttttgt tgcgaattta gttttcagtt ggtgaatttt 60gtaggtcaat ttagattatt atggcagttg ctttcgttat gatctgtatc attttcccat 120cctgtatcta cccagtgtat tatgttgagc tgtaagttac ttgaatgtga agcatgtaag 180cattcgaatt cattgtttaa ctcctaattc tagttccaca tgttatgttt tt 23224882DNAGlycine max 248ccatcctgta tctacccagt gtattatgtt gagctgtaag ttacttgaat gtgaagcatg 60taagcattcg aattcattgt tt 82249406DNAGlycine maxunsure at all n locations 249acgcccacgc gtccgtacgg ctgcgagaag acgacagaag ggggtgttgg atgaggcgaa 60actcgagagt gtaaggtttt gcatttcatt tgacgaagag tgagagagtc ttatctgtcg 120tctctgatct ctgatcgcat cttcattccg aaaatggctg tttcggctat cactggagcg 180aggctaactc tagggatgtc tctttcctct tccacgcgat cacgaaccgt cgcaatggcc 240gtatctatcg accccaagac cgataacana ctcactctta ccaagtccga ggaagcttcc 300gctgcggcca aagagctgat gcctggaggc gtgaactccc cagttcgtgc cttcanatcc 360gtgggtggtc anacaattgt gattgattca gtcaaagggt ctcgta 406250305DNAGlycine max 250cccacgcgtc cgtacggctg cgagaagacg acagaagggg gagagtgtaa ggttttgcat 60ttcatttgac gaagagtgag agagtcttat ctgtcgtctc tgatctctga tcgcatcttc 120attccgaaaa tggctgtttc ggctatcact ggagcgaggc taactctagg gatgtctctt 180tcctcttcca cgcgatcacg aaccgtcgca atggccgtat ctatcgaccc caagaccgat 240aacaaactca ctcttaccaa gtccgaggaa gctttcgctg cggccaagga gctgatgcct 300ggagg 305251296DNAGlycine max 251gaaactcgag agtgtaaggt tttgcatttc atttgacgaa gagtgagaga gtcttatctg 60tcgtctctga tctctgatcg catcttcatt ccgaaaatgg ctgtttcggc tatcactgga 120gcgaggctaa ctctagggat gtctctttcc tcttccacgc gatcaacaac acaagcaatg 180gccgtatcta tcgaccccaa gaccgataac aaactcactc ttaccaagtc cgaggaagct 240ttcgctgcgg ccaaggagct gatgcctgga ggcgtgaact ccccagttcg tgcctt 296252266DNAGlycine max 252ctgcgagaag acgacagaag ggggagagtg taaggttttg catttcattt gacgaagagt 60gagagagtct tatctgtcgt ctctgatctc tgatcgcatc ttcattccga aaatggctgt 120ttcggctatc actggagcga ggctaactct agggatgtct ctttcctctt ccacgcgatc 180acgaaccgtc gcaatggccg tatctatcga ccccaagacc gataacaaac tcactcttac 240caagtccgag gaagctttcg ctgcgg 266253293DNAGlycine maxunsure at all n locations 253ggttttgcat ttcatttgac gaagagtgag agagtcttat ctgtcgtctc tgatctctga 60tcgcatcttc attccgaaaa tggtgtttcg gctatcactg gagcgaggta actctaggga 120tgtctctttc ctcttccacg cgatcacgaa ctgaagcaat ggccgtatct atcgacccca 180agaccgataa caaacncatc ttaccaagtt cgaggaagtt tcgctgcggc caaggagtga 240tgctggaggc gtgaactccc cagttcgtgc cttcaaatcc gtgggtggtc aac 293254273DNAGlycine max 254gttggagagg cgaaactcga gagtgtaagg ttttgcattt catttgacga agagtgagag 60agtcttatct gtcgtctctg atctctgatc gcatcttcat tccgaaaatg gctgtttcgg 120ctatcactgg agcgaggcta actctaggga tgtctctttc ctcttccacg cgatcacgaa 180tccccgcaat ggccgtatct atcgacccca agaccgataa caaactcact cttaccaagt 240ccgaggaagc tttcgctgcg gccaaggagc tga 273255267DNAGlycine maxunsure at all n locations 255gggcgaaact cgagagtgta aggttttgca tttcatttga cgaagagtga gagagtctta 60tctgtcncct ctgatctctg atcgnatctn cattccgaan atggctgttt cggctatcac 120tggnncgagg ctaactctan ggatgtcnct ntnctcttcc angngatcac gcnntnnncg 180naanggacgn anctatcgac cccaagacng ataacaaatn actctnacca ngtccgngga 240agctttcgct gcggccaagg agntnat 267256254DNAGlycine max 256ggcgaaactc gagagtgtaa ggttttgcat ttcatttgac gaagagtgag agagtcttat 60ctgtcgtctc tgatctctga tcgcatcttc attccgaaaa tggctgtttc ggctatcact 120ggagcgaggc taactctagg gatgtctctt tcctcttcca cgcgatcacg aacccatgca 180atggccgtat ctatcgaccc caagaccgat aacaaactca ctcttaccaa gtccgaggaa 240gctttcgctg cggc 254257254DNAGlycine maxunsure at all n locations 257gttggatgag gcgaaactcg agagtgtaag gttttgcatt tcatttgacg aagagtgaga 60gagtcttatc tgtcgtctct gatctctgat cgcatcttca ttccgaaaat ggctgattcg 120gctatcactg gagcgccgtt aactctaggg atgtcttctt cctcgtgcag gcgacctcga 180acgctggnaa tggccgtatc tatcgacccc aagaccgata acaaactcac tcttaccaag 240tccgaggaag cttt 254258270DNAGlycine maxunsure at all n locations 258aggttttgca tttcatttga cgaagagtga gagagtctta tctgtcgnnt ctgatntntg 60atcgcatctt cattccgaaa atggcngttt cggctatcac tggagcgagg ctaagtntag 120ggatgtctct ttacctnttc cacgcgatca cgaaccacac gcaatggccg tatctatcga 180cccnaagacc gctaacaaan tcantctnac caagttccga ggaagntttg gnngcgggcc 240aagggagtga tgcctggagg cgtgaactcc 270259165DNAGlycine max 259ggcgaaactc gagagtgtaa ggttttgcat ttcatttgac gaagagtgag agagtcttat 60ctgtcgtctc tgatctctga tcgcatcttc attccgaaaa tggctgtttc ggctatcact 120ggagcgaggc taactctagg gatgtctctt tcctcttcca cacaa 165260161DNAGlycine maxunsure at all n locations 260cgaaactcga gagtgtaagg ttttgcattt catttgacga agagtgagan agtcttatct 60gtcgtctctg atctctgatc gcatcttcat tcccgaaaat ggctgtttcg gctatcactg 120gagcgaggct aactctaggg atgtctcttt cctcttccac a 161261153DNAGlycine max 261aaggttttgc atttcatttg acgaagagtg agagagtctt atctgtcgtc tctgatctct 60gatcgcatct tcattccgaa aatggctgtt tcggctatca ctggagcgag gctaactcta 120gggatgtctc tttcctcttc cacacaacat acg 153262241DNAGlycine max 262cttcatttga cgaagagtga gagagtctta tctgtcgtct ctgatctctg atcgcatctt 60cattccgaaa atggctgttt cggctatcag tggagcgagg ctaactctag ggatgtctct 120ttcctgttcc acgcgatgta taagatgatg gatggccgca tctatcgacc tctagacagc 180taagatactc agtcttagga ggtccgagga agctttcgct gtggccaagg attgatgtcc 240a 241263130DNAGlycine maxunsure at all n locations 263gcgaaactcg agagtgtaag gttttgcatn tcatttgacg aagagtgaga gagtcttatc 60tgtcgnntct gatctctgat cgcatcttca ttccgaaaat ggctgtttcg gctatcactg 120gagcgaggct 130264169DNAGlycine max 264cgctcgagcg aatcggctca cggctcgagg ttttgcattt actttgacga agagtgacga 60gagtcttatc tgtcgtctct gatctctgat cgcatcttca ttccgaaaat ggctgtttcg 120gctatcactg gagcgaggct aactctaggg atgtctcttt cctcttcca 169265181DNAGlycine maxunsure at all n locations 265gcgaaactcg anagtgtaag gnttngcatt ncanttgacg aagagtgaga gagtctnatc 60tgtcgngctc tgatntnnga tcgcatcntc attccganaa tggctgtttc ggctatcact 120ggagcgaggc taactctagg gangtctctn ncctcttcca cacaacatac gagnntcntc 180g 181266342DNAGlycine maxunsure at all n locations 266anacactgnt aaagtgaaga nggtgaatgg agatgtgtct gagaacaaca aaggaggnag 60caaaccttca gcagaaatag atcttccaga tgctgaagtt ggaaaagttc gcttgcgatt 120tgcacctgaa ccaagtggtt atcttcatat tggacactca aaagcagctt tgttgaacaa 180tattttgctg agcgatacca gggtcaggtt attgtncgnt ctgatgatan caatcctgct 240aaagagagca atgaatttgt ggacaacctg attaaagata ttgatacatt gggcatcana 300tatgaacaaa ttacatatac atcagattac ttccctgagt tg 342267290DNAGlycine max 267agctgccgga gataaagcta caacatatac taaaaggata tggcttgacc ttgctgatgc 60agtgtcttta tcagcaggtg aggaagtaac attgatggat tggggaaatg ccatagtgaa 120ggaaatagag aaggaccaag atggaaatat catagggttg agtggtgttt tgcatctaga 180aggatctgtg aagaccacaa aattgaaact cacttggcta cctgagatag atgaactagt 240tagcctgaca ttagtggagt ttgattatct aattacaaag aaaaagcttg 290268248DNAGlycine max 268tcggaattca gcgcgaggga tagcaatcct gctaaagtaa gcaatgaatt tgtggacaac 60cttattaaag atggtgatac attgggtatc aaatatgaac aaatgacata tacgtcagag 120tacttccctg agttgatgga gatggctgaa aaattaattc gccagggtaa agcatatgtt 180gatgacacac cacgtgaaca aatgcaaaaa gagagattgg atggcataga ttctaaatgc 240agaaataa 248269258DNAGlycine max 269ggcattgttg tgtggcggca cgccatggtc gaaggttact atttcaccat tttccaccac 60tcccacaccc ctcgcacctt cttcttccaa cgacgccgtt tctcagtctc tgctgctttc 120tccgaacaac aaccaccgcc acccgttcgc gttcgtttcg ctccttctcc caccggaaac 180ctccacgtcg gcggtgcccg aacggccctc ttcaactact tgttcgcaag gtccaaaggt 240gggaaatttg tgctgaga 258270267DNAGlycine max 270actgagtaga tggagatgga tgaaaaatta gttcgccagg gaaaagcata tgttgatgac 60atagcacgtg aacaaatgca aaaagagaga atggatggca tagattctaa atgcagaaat 120aatagtgtag aggagaatct aaaattgtgg aaggaaatgt tggcaggaac agagaggggg 180ttgcagtgtt gtgtccgtgg caagttggat atgcaggacc caaacaaatc acttagagat 240cctgtttatt atcgttgcaa tccaatg 267271245DNAGlycine max 271tgatgcacga tttcctacag tgcaaggaat tgtgcgtaga ggtttgaaaa ttgaagccct 60gatacagttt attgttgagc agggggcgtc caaaaatctc aatctcatgg aatgggacaa 120gctctggacc attaataaga agattattga ccctgtctgt cctagacaca ctgctgtcat 180tgcagacaga cgtgttttgt tgactctcac tgatggtcct gagtatcctt ttgtccgcat 240catac 245272280DNAGlycine max 272attgcaggaa cagagagggg cttgcagtgt tgtgtccgtg gcaagttgga tatgcaggac 60ccaaacaaat cacttagaga tcctgtttat tatcgttgca atccaatgcc ccatcataga 120attggatcca agtataaagt gtatccaact tatgattttg cttgtccata tgttgattct 180atagaaggaa tcacgcatgc ccttcgatct agtgaatacc atgatcgcaa tgcccagtat 240tactggattc aagaggacat gggtcttaga aaagttctta 280273276DNAGlycine max 273aggttgagtg gtgttttgca tcttgaagga tctgtgaaga ccacaaaatt gaaactcact 60tggctacctg agatagatga actagttagc ctgacattag tggagtttga ttatctaatt 120acaaagaaaa agcttgaaga agggaggatt tcattgatgt ggttaaccca tgtaccaaaa 180aggagacttt agcttatgga gactccaaca tgcgaaatct tcagcgtgga gatttattgc 240aactggagag aaagggatat ttcaggtgtg atttac 276274283DNAGlycine max 274agcaggtatt cgtgctgagt cagattctag agataattat tctcctggat ggaagtattc 60caactgggaa atgaaagggg ttcctctaag aattgaaatt gggccaaagg atttagcaaa 120taagcaggtc atcaactttg ccagtgtttt atcaattctc atatttgtca ttttgcttcc 180acactgttag tttttcagtg aacaccaaat aaatctcttt gaattttgca taggttcgca 240ctgttcgacg tgataatggt gcaaagatag acattgctag tgc 283275403DNAGlycine max 275caaaaccatt tgcgttgtcg cagtcgcagt caaaggccaa ggcaaaaccc taaattgtct 60cacactttcg tcggaatccg cttttggctt tttccgtgac aagatgccgg cgaaggacga 120cggctccgac aaggagaagt gccttgatct ctttctgaaa atcggcttag acgagcgcac 180cgctaaaaac accgtcgcaa acaacaaagt caccgccaat cttactgcag tcatctacga 240ggccggtgtt attgatggat gcagccgagc ggttggaaat cttctttaca cggttgcaac 300gaagtaccct gcaaatgcct tgccacatcg cccaacattg ctacagtaca ttgtctcgtt 360aaggtgaaaa caactgcaca gttagatgca gcattatcat ttc 403276445DNAGlycine maxunsure at all n locations 276gagaaaatgg cgctgctgtg angcggttgc catggnacga aggtnaatag tgnctctaca 60tgttnnaatc aatcntaaca ccccnaggna cntnnttatt cnaangacgc aagtttctna 120atctctgatg tctttagaac aacgnaacat ccgctcgnag tcgttttgct ncttctacaa 180cggaaacctt acatatcggc atgttccacg aacgggccct cttnaactac ttgttcgnaa 240ggtccaaang tggaaaattt gtgctgaata attgaggaca ctgacttgga naggtccagt 300agggagttat gaggaggcca atgctcaaag atctttcttg gcttggactt gattgggatn 360aaggncctgg tgttgaacgg gattatggcc ttatangcag tctgagagga attcttatcc 420aaccaatntc nggaaaacct acanc 445277277DNAGlycine maxunsure at all n locations 277gtttattatc gttgcaatcc aatgcnccat catagaattg gatccaagta taaagtgtat 60ccaacttatg attttgcttg tccatatgtt gattctatag aaggaatcac gcatgccctt 120cgatctagtg aancccatga ttgcaatgcc cagtattact ggattcaaga ggacatgggt 180cttagaaaag ttcttatcta cgaatttagc cggtncgaat atggtctaca ctcttctgag 240caaacgaaag cttttgtggt ttgtacaaaa tgggaaa 277278255DNAGlycine max 278agattctaga gataattatt ctcctggatg gaagtattct aattgggaaa tgaaaggtgt 60tcctctaaga attgaaattg ggccaaagga tttagcaaat aagcaggttc gtgctgttcg 120acgtgataat ggagcaaaga tagcattgct agtgctgatt tggttgtgga aataaaaaag 180ttgcttgata ctattcaaca gaacctgttt gatgttgcaa aacaaaaacg agatgaatgc 240attcagatca tacac 255279258DNAGlycine max 279agattctaga gataattatt ctcctggatg gaagtattct aattgggaaa tgaaaggtgt 60tcctctaaga attgaaattg ggccaaagga tttagcaaat aagcaggttc gtgctgttcg 120acgtgataat ggagcaaaga tagacatgct agtgctgatt tggttgtgga aataaaaaag 180ttgcttgata ctattcaaca gaacctgttt gatgttgcaa aacaaaaacg agatgaatgc 240attcagatca tacacact 258280265DNAGlycine max 280agattctaga gataattatt ctcctggatg gaagtattct aattgggaaa tgaaaggtgt 60tcctctaaga attgaaattg ggccaaagga tttagcaaat aagcaggttc gtgctgttcg 120acgtgataat ggagcaaaga tagacattgc agtgctgatt tggttgtgga aataaaaaag 180ttgcttgata ctattcaaca gaacctgttt gatgttgcaa aacaaaaacg agatgaatgc 240attcagatca tacacacttg ggatg 265281264DNAGlycine maxunsure at all n locations 281tcctgctaaa gaaagcaatg aatttgtgga caaccttatt aaagatattg atacattggg 60tatcaaatat gaacaaatta catatacgtc agattacttc cctgagttga tggagatggc 120tgaaaaatta attcgccagg gtaaagcata tgttgatgac acaccacgtg aacaaatgcn 180aaaagagaga atggatggca tagattctaa atgcagaaat aatagtgtag aggagaatct 240aaaattgtgg aaggnaatga ttgc 264282263DNAGlycine max 282cctgattaaa gatattgata cattgggcat caaatatgaa caaattacat atacatcaga 60ttacttccct gagttgatgg aaatggctga aaaattaatt cgcgagggta aaacatatgt 120tgatgacact ccacgtgaac aaatgcaaaa agagagaatg gatggcatag aatctaaatg 180cagaaataat atagtagagg agaatctaaa actgtggaag gaaatgattg caggaacaga 240gaggggattg cagtgttgtg tcc 263283267DNAGlycine max 283ttgggcatca aatatgaaca aattacatat acatcagatt acttccctga gttgatggaa 60atggctgaaa aattaattcg cgagggtaaa acatatgttg atgacactcc acgtgaacaa 120atgcaacaag agagaatgga tggcatagaa tctaaatgca gaaataatat agtagaggag 180aatctaaaac tgtggaagga aatgattgca ggaacagaga ggggattgca gtgttgtgtc 240cgtggcaagt tggatatgca ggaccca 267284269DNAGlycine max 284atgggagttc agcaaaccca ctccattcat caggagtcgc gagtttcttt ggcaagaagg 60gcacactgct tttgcaacaa aggatgaagc agatgcagag gttcttgaga ttctggaatt 120atataggcgt atatacgaag agatttggca gttcctgtca taaagggtaa gaaaagtgag 180cttgagaagt ttgctggtgg actctacact accagtgttg aggcatttat tccaaacact 240ggtcgtggta tccaaggtgc aacttctca 269285422DNAGlycine max 285gtccaaacgg cagcgagaag acgacagaag gggtcagatg ggagttcagc aaccccactc 60cattcatcag gagtcgtgag tttctttggc aagaagggca cactgctttt gcttcaaagg 120aggaagcaga tgcagaggtt cttgagattc tggaattata taggcgtata tacgaagagt 180atttggcagt tcctgtcata aagggtaaga aaagtgagct tgagaagttt gctggtggac 240tctacactac tagtgttgag gcatttattc caaacactgg tcgtggtata caaggtgcaa 300cttctcattg tttgggccaa aattttgcta aaatgtttga gataaacttt gaaaatgaaa 360agggagagag agcaatggtc tggcagaatt catgggccta

tagtactcga actatcggtg 420tc 422286240DNAGlycine max 286aaattatata ggcgtatata cgaagagtat ttggcagttc ctgtcataaa gggtaagaaa 60agtgagcttg agaagtttgc tggtggactc tacactacca gtgttgaggc atttattcca 120aacactggtg tggtatccaa ggtgcaactt ctcattgttt gggccaaaat tttgctaaaa 180tgtttgagat aaactttgaa aatgaaaagg gagagaaagc aatggtctgg cagaattcat 240287378DNAGlycine max 287ggaggctaca atttttgagc tacgttatcg aacaaatgtg ggtgagttgc ttgggcgtgt 60gcgcaaagag ctgccatggg gtgatgcaaa agttgccaag caacttgttg atgcgcaact 120atatgaacta cttggtgatc ggacagcagc agatgatgaa aagccttcta gaaagaagaa 180ggagaaacct gctaaagtag aggataaggc agctcctgtt tctacccctg aaaagtcacc 240tgaagaagac gttaatccat ttttaatatt ccctaatcca gaggaaaatt tcaaggtgca 300tactgaagtg ccttttagtg atggtagtat tttgagatgt tgcaatacaa gagatctgct 360tgacaaacac ttaaaagc 378288269DNAGlycine max 288aacaaatgca aaaagagaga atggatggca tagaatctaa atgcagaaat aatatagtag 60aggagaatct aaaactgtgg aaggaaatga ttgcaggaac agagagggga ttgcagtgtt 120gtgtccgtgg caagttggat atgcaggacc caaacaaatc acttagagat cctgtatatt 180atcgttgcaa tccaatgccc catcatagaa ttggatccaa gtataaagtg tatccaactt 240atgatttcgc ttgtccatat gttgatgct 269289258DNAGlycine max 289aacaaatgca aaaagagaga atggatggca tagaatctaa atgcagaaat aatatagtag 60aggagaatct aaaactgtgg aaggaaatga ttgcaggaac agagagggga ttgcagtgtt 120gtgtccgtgg caagttggat atgcaggacc caaacaaatc acttagagat cctgtatatt 180atcgttgcaa tccaatgccc catcatagaa ttggatccaa gtataaagtg tatccaactt 240atgatttcgc ttgtccat 258290251DNAGlycine maxunsure at all n locations 290aggcgatctc ggttgggaag cggggaagat ggggaagctt gtaattaagc atttggctgc 60caacncggtg cagaagaatg gttgttgtta acaggactga agagaaagtt aatgccattc 120ggaaagagtt gaaggatgtt gagattgtat ttagaccatt ttcagatatg ctggcgtgtg 180ctgctgaagc tgatgtgatc ttcaccagca cagcgtctga atcaccatgt tctctaaaca 240gaatgtgcag a 251291240DNAGlycine max 291atttgcatag ggctgaacat tcacactgct cccgttgaga tgcgtgagaa gcttgcaatt 60ccagaatccc attgggctca ggctattaag gacctttgcg ctttgaacca tatcgaagaa 120gccgcggttc tcagcacgtg taaccgcatg gagatctatg ttgtggctct ttcccagcac 180cgtggtgtta aggaagttac tgattggatg tctaaggtga gcgggatttc aatacctgag 240292275DNAGlycine maxunsure at all n locations 292aggaagcagc tgttctgagc acctgcaaca gaatggaaat atatgttgtt gctctgtcca 60agcaccgtgg tgttaaagaa gtcactgaat ggatgtccaa aacangtggg attccagttg 120cagatctttg ccagcatcag tttctgctat acaacaaaga tgccacacaa cacctttttg 180aagtatctgc aggtcttgat tctctagtgt tgggagaagg tcaatccttg cccaggtgan 240gcaggttgtc aatttggaca aggnttaang ncttc 275293276DNAGlycine maxunsure at all n locations 293ggtaagaact tgagacaaaa cattgctgct ggtgcagtan ncnnnnagtt catcaactgt 60antncnggga cntnattnag gctaccngaa gnctcacatg ncatgcaagg ntgttggtca 120ttggagctgg gnagatcgga agcttgtgat caagcatttn gtggcaaaag ggtgcacaaa 180gatggtggtt gtcatagagt gangagagag ttgccgcgat ccgtgaagaa atcaagatgt 240tgagataatc tacaagccac tctcggagat gctcac 276294271DNAGlycine max 294ctcgagcgga ataagctact tcatggtccc atgcagcacc taaggtgtga tgggaacaat 60gatagtagtc tgagtgaagt acttgagaat atgcgcgccc ttaacagaat gtatgatctt 120gagacagaaa cttccttgat cgaagaaaag atcagagtca agatggaacg ggttcagaag 180tagattcttc ttcaattggt ttagttttac ttgattactg tgggggctgc aatcctcgcc 240attttgtaca ctacagtagt tgattgaggc c 271295130DNAGlycine max 295ggcaatcatt gctgaagaat ctaagcaatt tgaagcttgg agggactcgc tggaaactgt 60tcctactatt aagaaattga gggcttatgc tgaaagaatc aggcttgctg agcttgagaa 120gtgcttaggt 130296426DNAGlycine max 296cccacgcgtc cgaacatttg gtggcaaaag gttgcaaaaa gatggtggtt gtcaatagaa 60ctgatgagag agttgctgca atacgtgaag aactgaagga tattgagatt atctacaaac 120ccctttcaga aatgctcacc tgtgctggcg aagcagattt agttttcacc agtactgcat 180cagaaaaccc attattcttg aaagaacatg tcaaggacct tcctcctgca agtcaagaag 240ttggaggccg tcgctttttc attgatatct ctgttccccg gaatgtgggt tcatgtgtct 300cagaccttga gtctgtgcga gtttacaatg ttgacgacct taaagaggtt gtggctgcca 360ataaagagga tcgcctaaga aaagcaatgg acgcacaggc aatcattgct gaaaaatcta 420agcaat 426297271DNAGlycine max 297aggataggct aagaagagcc atggaggctc aagcaatcat tggtgaagaa tcaaaacaat 60ttgaggcttg gagagactca ttggaaactg ttcctaccat taaaaagttg agggcatatg 120ctgaaagaat aaggcttgct gagcttgaga agtgcctagg taagatgggt gatgatatca 180acaagaagac acaaagagct gtggatgatc ttagcagggg tatagtgaat aagttgcttc 240atgggccaat gcaacacttg aggtgtgatg g 271298266DNAGlycine max 298agaaaagcca tggaggctca agcaatcatt ggtgaagaat caaaacaatt tgaggcttgg 60agagactcat tggaaactgt tcctaccatt aaaaagttga gggcatatgc tgaaagaata 120aggcttgctg agcttgagaa gtgcctaggt aagatgggtg atgatatcaa caagaagaca 180caaagagctg tggatgatct tagcaggggt atagtgaata agttggcttc atgggccaat 240gcaacacttg agtgtgatgg cagtga 266299289DNAGlycine max 299cacaattctc ccttcaaagt ttcaatggct gtttcaacca gcttctcggg tgtaaagttg 60gaggctttgt tgctgaaatg tggttcctcc aatgctgcca ccaccaccac tcatatatca 120tgttttggca aaaacagaaa gacacttgtt cagagtcaga gaggggctat tcgttgtgag 180gcttcttctg cttctgatgt tgtggctgat gccaccaaga aagctgctag tgtctctgct 240cttgagcagc ttaagacctc tgcagctgat aggtatacaa aggaaagga 289300289DNAGlycine maxunsure at all n locations 300cacaattctc ccttcanagt ttcaatggct gtttcaacca gcttctcggg tgtaaagttg 60gaggctttgt tgctganatg tggttcctcc aatgctgcca ccaccaccac tcatatatca 120tgttttggca aaaacagaaa gacacttgtt cagagtcaga gaggggctat tcgttgtgag 180gcttctnctg cttctgatgt tgtggctgat gccaccaaga aagctgctan tgtctctgct 240cttgagcagc ttaagacctc tgcagctgat aggtatacna aggaaagga 289301266DNAGlycine max 301cagggcttga ctcacttgtt cttggggaag gtcaaattct tgctcaggtg aagcaggttg 60tgaaagctgg acagggagtg cctggttttg ataagaaaat cagtggtttg ttcaagcagg 120cgatatcggt tgggaagcgg gttagaaccg agactaacat ttcatctgga tcagtttctg 180taagctcggc tgctgtggag cttgcactga tgaagctacc ggaaattacc tttgctgatt 240ctggagtgtt ggtggttggt gctggg 266302275DNAGlycine max 302cgcgcacatc tatttgaagt ggcgtcaggg cttgactcac ttgttcttgg ggaaggtcaa 60attcttgctc aggtgaagca ggttgtgaaa gctggacagg gagtgcctgg ttttgataag 120aaaatcagtg gtttgttcaa gcaggcgata tcggttggga agcgggttag aaccgagact 180aacatttcat ctggatcagt ttctgtaagc tcggctgctg tggagctgca ctgatgaagc 240taccggattc ctcctttgct gattctggag tgttg 275303288DNAGlycine max 303cttgagcagc ttaagacctc tgcagctgat aggtatacaa aggaaaggag cagcatcatg 60gttattggac tgagtgtgca tagtacacct gtggaaatgc gtgaaaaact tgccatacca 120gaagcagaat ggccaagagc cattgcggag tttgtagtct gaatcatatt gaggaagcag 180ctgttctgag cacctgcaac agaatggaga tatatgttgt tgctctgtcc aagcaccgcg 240gtgtcaaaga agtcactgaa tggatgtcca aaacaagtgg gatcccgg 288304299DNAGlycine max 304agtgtgcata gtacacctgt ggaaatgcgt gaaaaacttg ccataccaga agcagaatgg 60ccaagagcca ttgcggagtt tgtagtctga atcatattga ggaagcagct gttctgagca 120cctgcaacag aatggagata tatgttgttg ctcttccaag caccgcgttg tcaaagaagt 180cactgaatgg atgtccaaaa caagtgggat cccggttgca gacctttgcc agcatcagtt 240tctgctatac aacaaagatg cgacacagca cctttttgaa gtatctgctg gtcttgatt 299305260DNAGlycine maxunsure at all n locations 305gagcagcatc atggttattg gactgagtgt gcatagtaca cctgtggaaa tgcgtgaaaa 60acttgccata ccagaagcag aatggccaag agccattgcg gagtttgtag tctgaatcat 120attgaggaag cagcngttct gagcacctgc aacagaatgg agatatatgt ngttgctctg 180tccangcacc gcggtgtcaa agnagtcact gaatggntgt ccaaaacaag tnggntcccg 240gttgcagact ttgccagcat 260306440DNAGlycine max 306gggttctcct gaatccgcaa tggccgtttc aaccactttc tccggtgcca aattggaggg 60gctattgctc aaatgttctt cctcctcttc ctcaccaccg ccttcaaggt catcattcac 120cacttttccc ggccaaaaca gaagaaccct cattcagaga ggggttattc gctgcgacgc 180tcagccctct gatgcatcat ctgttgctcc aaataatgcc accgctctct ccgctcttga 240gcagctcaag acttctgcag ctgatagata tacaaaggaa agaagcagca ttatcgccat 300tgggctcagt gtgcacactg cacctgtgga aatgcgtgaa aaacttgcca ttccagaagc 360agaatggcct agagctattg cagagctgtg tagtctgaat catatttgag aagcagctgt 420tctgagtacc ctgcatcgaa 440307272DNAGlycine max 307ctgaaatcaa ggttgttgct ggtgaccctt ataactcaga cccacaagat ccagaattca 60tgggtgttga agtcagagag cgtgtacttc caaggagagg aactttctgt tgtcttgacc 120aaaattaaca tggttgattt gcattgggag ctacagaaga tagagtgtgt ggaacaattg 180acattgagaa agccctgact gagggtgtca aggcatttga gcctggacta tggctaaagc 240taatagggga atctatatgt tgatgaagtt aa 272308254DNAGlycine max 308gtcttacaac ggctttagag ttggactaaa tgcggagaaa agtggtgacg ttggacgtat 60aatgattgtt gcaatcactg atggcagagc caatatatca ttgaaaaggt caactgaccc 120tgaagctgcc gcagctactg atgccccaaa accttcagca caagaattga aggatgaaat 180tcttgaggtg gccggaaaga tatataaagc aggaatgtct ctccttgtca tcgacactga 240aaataagttt gtct 254309253DNAGlycine max 309actttctgtt gtcttgacca aaattaacat ggttgatttg ccattgggag ctacagaaga 60tagagtgtgt ggaacgattg acattgagaa agccctgact gagggtgtca aggcatttga 120gcctggacta ctggctaaag ctaatagggg aatcttatat gttgatgaag ttaatctttt 180ggatgatcac ttggtggatg tgttgttgga ttctgctgcg gatggaacac agtagagaga 240gagggaattt cta 253310253DNAGlycine max 310tgttactctt aacagagaac aattaaaata cctggttatt gaagctttac ggggcggttg 60ccagggacat agagctgatc tatttgctgc ccgtgttgca aagtgcttag ctgctttgga 120gggacgtgaa aaggtttatg tggatgacct aaaaaaagct gtagaattgg tcattctacc 180ccggtcaatc gttactgaga acccaccaga tcaacaaaac cagcctcctc cccctccgcc 240tcctccacaa aat 253311162DNAGlycine max 311gcatgatgat ctccacatgt ctgtctgtca actaaaacac tattgcgttt catgatatat 60caaattgtga acatgctatg tgttaatgtt tctttaaagc ataatccata gccccttatg 120tttaatcaaa ccaaaattat gccctagttt tttttttttt gg 162312232DNAGlycine max 312aaaaaagaac agagagagaa gaatgaaatc tatctatctt cttatccgaa gtctgggagg 60ccaataggaa gcacgccagc tgctacgaat ggtgaataaa agacaaaaga aacaaactgc 120tacatagcat acagtctgtc ttctcttctc ttctccggtt atggcgtccg ccttgggcac 180ttcttcaatt gcggttctgc cttcgcgcta cttctcttct tcttcttcca ag 232313262DNAGlycine maxunsure at all n locations 313cacttaatcc aggctcagaa gattgctttt aacgagagcc agangccggt gtacccattt 60tctgctatag tgggacacga tgagatgaag ctttgccttc tcctaaatgt aattnatccc 120aagattggag gtgtaatgat catgggggac agaggaacgg ggaaatctac aactgttaga 180tcattggtag atttgcttcc tgaaatcaag gttgttgctg gtgaccatat attcagaccc 240agaggatcca gattcatggg tg 262314280DNAGlycine maxunsure at all n locations 314actctctcta acttcagggc agagctatgg gcggaaattt tatggaggaa ttggaattca 60tggcatcaag ggaaggtctc agctctcagt tgccaatgtt gccactgaag ttaactctgt 120agaacaggcc caaagtattg cttctaaaga aagccagagg ccagtatacc cattttctgc 180catagtngga caagatgaga tgaagctttg tcttctcctt aatgtgattg atcctaagat 240tggaggtgta atgatcaggg ggataggggc acagggaaat 280315238DNAGlycine max 315ttttgctcgg aatttcctgt gtagaaggaa ctcatgaatc ttattgatgt ttaacgacaa 60tgaaaatctc cacagaaaag gtaaaatgta aataatgaag tagcattata ctcatggaat 120accacagaat acaaaccgtg ttacatctat gatcctcagc tgaatacctc ataaaatttc 180tcagtgacaa gtaaacctga gtctatagac tccaagggat cctttctaag acggtgtc 238316273DNAGlycine max 316ttagggaagg gctcagctct cggttaccaa tgttgccact gaagttaact ctgtagaaca 60ggctcagagt attgcttcta aagaaagcca gaggccagta tacccatttt ctgccatagt 120tggacaagat gagatgaagc tttgtcttct ccttaatgtg attgatccta agattggagg 180tgtaatgatc atgggggata ggggcacagg gaaatctaca acggtcaggt cattggttga 240tttacttccc gaaatcaagg ttgttgctgg tga 273317283DNAGlycine max 317agactcattg gatcggttga tgttgaggag tctgtgaaaa caggcacaac tgttttccag 60ccaggcttgc ttgcagaagc tcatagaggt gttttatatg ttgatgaaat taatcttttg 120gatgagggta tcagtaattt gctccttact gtattgagtg aaggagtaaa tactgttgaa 180agagagggga tcagtttcaa gcacccttgc aggccccttc tcattgccac ctataaccca 240gaagagggtg ctgttcgtga acatctgctg gaccgcattg cga 283318173DNAGlycine maxunsure at all n locations 318gctcgaggcg ccgntcanac gacgagccgc gagtgcgtgg cggcgtggga cgaggtggag 60gagctgagcg cggcggcgag ccacgccaaa tacaagctaa aggaaaagga ctccgacccg 120ctcgagacct actgcaagga caatccggag accattgagt gcaaaacttt cga 173319263DNAGlycine max 319aggaattccg agattcttac aaagccgagc aagagaagct ccaacaacaa attacatcag 60caaggagtgt tctttcttct gttcagattg atcaagatct caaggtgaaa atctccaagg 120tgtgtgctga gttgaatgtg gatggattaa gaggagacat agtaacaaat agagctgcaa 180aagctcttgc tgctctgaag gaaagagaca aagtaagtgc agaggatatt gctactgtca 240tccctaactg cttgagacac cgt 263320322DNAGlycine max 320atagctttgg gagcaaaaac tgcacaaagc tcctcagtgc cccccaagtt ttcctttcaa 60agcaattttg tgctttgctt tgaatgtctt ccttttcgat ccctacactt caatttgtag 120caagaggaat ttgttgtttc ctacttagca tgattattta tcaatggcgt ctttggtatc 180ttcagcattt actcttccaa gctctaaacc tgaccagctt caatcacttg ccccgaaaca 240tctttttcat cagtcattcc ttcccaagaa agccaattac aatggtagct caaaatcctc 300tctgaaaatt aaatgtgctg tc 322321410DNAGlycine maxunsure at all n locations 321cagtcattac tttgactcan accccgacta atctggntca gaatctaagg aaagatggga 60agaagcctag tgcatacatt gctgatacaa ccacagccaa tgctcaggta cgtacactnt 120ctgagacggt tagacttgac gcaagaacca agctgttgaa tccaaagtgg tatgaaggca 180tgttgtctac tggatatgag ggtgtacgcg agatcgagaa gagactcacc aatacagtgg 240ggtggagtgc aacttcaggc caagttgata actgggtgta tgaagaagcc aacacaactt 300tcattcaaga tgagcaaatg ctgaacaagc tcatgagcac taatccaaac tccttcagga 360aactggtgca gacattcttg gaagccaatg gacgtggtta ttgggaaact 410322324DNAGlycine max 322gaaaaataac acacatttga aactcaaact gaaatgggtg catagctttg gggcaaaaac 60tacacaaaac tcctcattgc ccccaagttt tttctttcaa agcaattttg cacttttttg 120ctttcattgt cttcaatttg tagtaagagg aaattgttgt ttcctactta gcttgattat 180tattatcaat ggcttcttta gtatcttcac aatttacact accaagttct aaacctgacc 240agcttcattc tcttgctcag aagcatcttt ttctccactc tttccttccc aagaaggcca 300attacaatgg tagcagctca aaat 324323340DNAGlycine maxunsure at all n locations 323gaagaagtaa tacatgacaa agaagctcaa tttagcagcc caaatctgaa cgttgcttac 60aaaatgaatg tccgagaata ccaaagtcta actccctatg ccacagcatt agaagaaaac 120tggggaaaac ctcctgggaa tctgaattca gatggagaga atctattggt atatgggaaa 180caatatggta atgtattcat aggtgttcaa cccacatttg gctatgaagg cgatcctatg 240cggttgcttt tctccaaatc tgcaagtcct catcatggat ttgcagcatn atactctttt 300gtttgagaaa ttttcaaagc tgaagcggtt cttcattttg 340324264DNAGlycine max 324ggcgaagaac agaatgaaga ggaagaacaa gaggatgaca aggatgaaga gaatgaacaa 60cagcaagaac aattacctga agagtttatc tttgatgctg aaggtggctt ggtagatgaa 120aaactcctct tctttgccca acaagcacag agacgccgtg ggagggctgg aagggcaaaa 180aatgttatat tttccgagga tagaggccga tacatcaagc ccatgcttcc aaagggccct 240gtaaagagat tagctgtaga tgca 264325246DNAGlycine max 325caaaatcaag aatcaggcga agaacagaat gaagaggaag aacaagagga tgacaaggat 60gaagagaatg aacaacagca agaacaatta cctgaagagt ttatctttga tgctgaaggt 120ggcttggtag atgaaaaact cctcttcttt gcccaacaag cacagagacg ccgtgggagg 180gctggaaggg caaaaaatgt tatattttcc gaggatagag gccgatacat caagcccatg 240cttcca 246326264DNAGlycine maxunsure at all n locations 326cnagagcaga gaagantcag agaatggcaa ctatgactgg cgtgagcctt tcatgcccca 60gggttttctt caacgcatca ggctcaccgc aaaacgcgca tgcttattgt attttgtcca 120gcagattcta tgacttgaca ggactgcaga atggaattct gaagcgaggg agagagattt 180tcctcactgg ttgctacctc cgaactccca ctggaggttc tggacattca cgtcttttgc 240caacagagta tcttgtgatt ctat 264327284DNAGlycine maxunsure at all n locations 327cagagaagaa tcagagaatg gcaactatga ctgnngtgag cntttcatgc cccagggttt 60tcttcaacgc atcaggctca ccgcaaaacg cgcatgctta ttgtattttg tccagcagat 120tctatgactt gacaggactg cagaatggaa ttctgaagcg agggagagag attttcctca 180cnngttgcta cctccgaact cccactggag gttctggaca ttcacgtctt ttgccaacag 240agtatcttgt gattctattg gatgaagact tccagaagga aatt 284328392DNAGlycine max 328ggccgataca tcaagcccat gcttccaaag ggccctgtaa agagattagc tgtagatgca 60acccttagag ctgctgcacc ttatcaaaaa ttgcgaaggg caaaagattc tggaaacaat 120agaaaggtat ttgtggagaa aacggacatg agggcaaaga gaatggcacg taaggcagga 180gcattggtga tatttgttgt tgatgcaagt ggaagcatgg cattgaacag gatgcagaat 240gcaaaaggtg cagcacttaa gcttctggct gaaagttata caagcaggga tcaggtatct 300ataattccat tccgtggaga tgcagctgaa gttctcctgc caccttctag atcaatttca 360atggcaagga aacgtcttga aaggcttcca tg 392329274DNAGlycine max 329gtggagaaaa cggacatgag ggcaaagaga atggcacgta aggcaggagc attggtgata 60tttgttgttg atgcaagtgg aagcatggca ttgaacagga tgcagaatgc aaaaggtgca 120gcacttaagc ttctggctga aagttataca agcagggatc aggtatctat aattccattc 180cgtggagatg cagctgaagt tctcctgcca ccttctagat caatttcaat ggcaaggaaa 240cgtcttgaaa ggcttccatg tggtggaggt cccc 274330247DNAGlycine max 330attagctgta gatgcaaccc ttagagctgc tgcaccttat caaaaattgc gaagggcaaa 60agattctgga aacaatagaa aggtatttgt ggagaaaacg gacatgaggg caaagagaat 120ggcacgtaag gcaggagcat tggtgatatt tgttgttgat gcaagtggaa gcatggcatt 180gaacaggatg cagaatgcaa aaggtgcagc acttaagctt

ctggctgaaa gttatacaag 240cagggat 247331292DNAGlycine maxunsure at all n locations 331tngagggcaa agagaatggc acgtaaggna ggancatcgg tgatatttgt ggttgatgca 60agtggaagca tggcattgaa caggatgcag aatgcaaaag gtgcagcact taagcttctg 120gctgaaagtt atacaagcag ggatcaggtc tctaaattcc attccgtgga gacgcagctg 180aagttcttct gccaccttct agatcaattg caancgnaag gaaacgtctt gagaggctcc 240atgtggtgga gggtccccac ttgctcaggt ctacaacggc tgttagagtt gg 292332378DNAGlycine max 332agacgggtgc gagaagacga cagaagggga taagtgccat aacacataaa cagaatggct 60tccacgtttg gcgcatcttc aattaccttc ctctcttcac gatactactc gtctcaggcc 120cttgccaccg attcaccctc tctaaccaca gtgcagatat ttgggcgcaa gttttgcgga 180ggaagaaatg gatttcacag cgtcaaggga aggtctctgt tcgcggttgc gagtgttctt 240gccactcaac ttaactctgc ataataggct cagaagattg cttttaccga gagccagagg 300tcagtgtacc cattttcggc tatagttgga caggatgaaa tgaagctttg ccttctccta 360aatgtgattg atcccaaa 378333277DNAGlycine max 333aaaaagaatg gcttccacgt ttggcgcatc ttcaattacc ttcctctctt cacgatacta 60ctcttcccaa tcccttgcca ccgattctcc ctctctaacc acagtgcaga tatttgggcg 120caagttttgc ggcggaggaa atggatttca cagcgtcaag ggaaggtctc tgttcccggt 180tgcgagtgtt cttgccactc aacttaactc tgcacaacag gctcagaaga ttgcttttac 240cgagagccag aggccagtgt acccatttcg gctatag 277334256DNAGlycine max 334taaaaagaat ggcttccacg tttggcgcat cttcaattac cttcctctct tcacgatact 60tctcttccca atcccttgcc accgattctc cctctctaac cacagtgcag atatttgggc 120gcaagttttg cggcggagga aatggatttc acagcgtcaa gggaaggtct ctgttcccgg 180ttgcgagtgt tcttgccact caacttaact ctgcacaaca ggctcagaag attgctttta 240ccgagagcca gaggcc 256335396DNAGlycine max 335ggcaactatg actggtgtga gcctttcatg ccccagggtt ttcttcaacg catcagcctc 60accgcaaaac gcgcatgctg taaagttctc acttccaccc agccaagcag tgcgaccggg 120tagtatcaag ttgggtcgcg tgatgaggat ccgacccgtt cgcgctgcgc ctgagcgcat 180atcggagaag gtggaggaga gcataaagaa cgcgcaggag gcgtgcgccg gcgatccgac 240gagcggcgag tgcgtggcgg cgtgggacga ggtggaggag ctgagcgcgg cggcgagcca 300cgccagggac aagcaaaagg aaaaggactc cgacccgctc gagaattact gcaaggacaa 360cccggagacc attgagtgca aaactttcga agactg 396336356DNAGlycine max 336gagaatggca actatgactg gtgtgagcct ttcatgcccc agggtggtct tcaacgcatg 60agcctcaccg cataacgcgc atgctgtaaa gttctcactt ccacccagcc aagcagtgcg 120accgggtagt atcaagttgg gtcgcgtgat gaggatccga cccgttcgcg ctgcgcctga 180gcgcatatcg gagaaggtgg aggagagcat aaagaacgcg caggaggcgt gcgccgacga 240tccgacgagc ggcgagtgcg tgacggcgtg ggacgaggtg gaggagctga gcgcggcggc 300tagccacgcc agggacacgc aaatggtaat ggacttcgac ccgctcgaga attact 356337273DNAGlycine max 337agaatggcaa ctatgactgg tgtgagcctt tcatgcccca gggttttctt caacgcatca 60gcctcaccgc aaaacgcgca tgctgtaaag ttctcacttc cacccagcca agcagtgcga 120ccgggtagta tcaagttggg tcgcgtgatg aggatccgac ccgttcgcgc tgcgcctgag 180cgcatatcgg agaaggtgga ggagagcata aagaacgcgc aggaggcgtg cgccggcgat 240ccgacgagcg gcgagtgcgt ggcggcgtgg gac 273338272DNAGlycine maxunsure at all n locations 338aagaatcaga gaatggcaac tatgactggt gtgagccttt catgccccag ggttttcttc 60aacgcatcag cctcaccgca aaacgcgcat gctgtaaagt tctcacttcc acccagccaa 120gcagtncgac cgggtagtat caagttgggt cgcgtgatga ggatccgacc cgttcgcgct 180gcgcctgagc gcatatcgga gaaggtggag gagagcataa agaacgcgca ggaggcgtgc 240gccggcgatc cgacgagcgg cgagtgcgtg gc 272339273DNAGlycine maxunsure at all n locations 339gaatcagaga atggcaacta tgactggtgt gagcctttca tgccccaggg ttttcttcaa 60cgcatcagcc tcaccgcaaa acgcgcatgc tgtaaagttc tcacttccac ccagccaagc 120agtccgaccg ggtagtatca agttgggtcg cgtgatgagg atccgacccg ttcgngtgcg 180cctgagcgca tatcggagaa ggtggaggag agcataaaga acgcgcagga ggcgtgcgcc 240ggcgatccga cgagcggcga gtgcgtggcg gcg 273340253DNAGlycine max 340cagagaatgg caactatgac tggtgtgagc ctttcatgcc ccagggtttt cttcaacgca 60tcagcctcac cgcaaaacgc gcatgctgta aagttctcac ttccacccag ccaagcagtg 120cgaccgggta gtatcaagtt gggtcgcgtg atgaggatcc gacccgttcg cgctgcgcct 180gagcgcatat cggagaaggt ggaggagagc ataaagaacg cgcaggaggc gtgcgccggc 240gatccgacga gcg 253341283DNAGlycine maxunsure at all n locations 341gtaactatga ctggtgtgag cctttcatgc cccagggttt tcttcaacgc atcagcctca 60ctgnaaaacg cgcatgatgt aaagttctca cttccacaca gcatagaagg tggatcgggt 120agtatcaagt tgggtcgcgt gatgaggatc cgagccgttc gcgctgcgcc tgagcgcata 180tcggagaagg tggaggagag catacagaac gcgcaggagg cgtgcgccgg cgatcagttg 240agcggcgagt gcgtggcggc gtgggacgat gtggaggagc tga 283342251DNAGlycine max 342gagaatggca actatgactg gtgtgagcct ttcatgcccc agggttttct tcaacgcatc 60agcctcaccg caaaacgcgc atgctgtaaa gttctcactt ccacccagcc aagcagtgag 120accgggtagt atcaagttgg gtcgcgtgat gaggatccga cccgttcgcg ctgcgcctga 180gcgcatatcg gagaaggtgg gagagcataa agaacgcgcg gaggctgcgc ggcgatccga 240cgagcggcga t 251343271DNAGlycine max 343aaaccccctc cagagaacaa gaatcaaaga atggcaacta tgactggtgt gagcctttca 60agccccaggg ttttcttcaa cgcatcaccc tcaccgcaaa acacgtacgc cgtaaagttc 120gcagttccac tcagccaagg gatgcgactt ggtagtgtca ggttgggtcg ggtgatgagg 180atccgacccg ttcgcgcagt ccagagcgca tttcggagaa ggtggaggag agcataaaga 240acgcgcagga ggcgtgcgcc ggcgacccga c 271344257DNAGlycine max 344gcctttcaag ccccagggtt ttcttcaacg catcaccctc accgcaaaac acgtacgccg 60taaagttcgc agttccactc agccaaggga tacgacttgg tagtgtcagg ttgggtcggg 120tgatgaggat ccgacccgtt cgcgcactcc agagcgcatt tcggagaagg tggaggagag 180cataaagaac gcgcaggagg cgtgcgccgg cgacccgacg agcggcgagt gcgtggcggc 240gtgggacgag gtggagg 257345281DNAGlycine maxunsure at all n locations 345gagaatggca actatgactg gtgtgagcct ttcatgcccc agggttttct tcaacgcatc 60agtctcaccg naaaacgcgc atgctgtaaa gttctcactt tcanacagcc aagaagacac 120aaagggtagt atcaagttgg gtcgcgtgat gaggatccga cccgttcgag ctgcgtctga 180gcgcatatcg gagaaggtgg aggagagctg aaggaacgcg caggaggcgt gcgccggcga 240tccgacgagc ggcgagtgcg tagcggcgtg ggacgaggtg g 281346249DNAGlycine max 346gagaatggca actatgactg gtgtgagcct ttcatgcccc agggttttct tcaacgcatc 60agcctcaccg caaaacgcgc atgctgtaaa gttctcactt ccagccagcc tatgagtctt 120accgggtagt agcaagttgg gtcgcgtgat gatgatccga cccgttcgcg ctgcgcctga 180gcgcatatcg gagaaggtgg aggagagcaa acagaacgcg ctaggaggcg tacgccggcg 240atccgacga 249347240DNAGlycine max 347cgtccgatag gatgcgagaa gacgacagaa ggggagagaa caagaatcaa agaatggcaa 60ctatgactgg tgtgagcctt tcaagcccca gggttttctt caacgcatca ccctcgccgc 120aaaacacgta cgccgtaaag ttcgcagttc cactcagcca agggactcga cttggtagtg 180tcaggttggg tcgggtgatg aggatgcgag ccgttcgcgc agctccagag cgcagttcgg 24034891DNAGlycine max 348gagaatggga actatgactg gtgtgagcgt ttcatgcgcc agggttttct gcaacgcatc 60agcgtcaggg caaaacgcgc atagtgtaaa g 91349119DNAGlycine max 349ctcgagccga gagaatggca actatgactg gtgtgagcct ttcatgcccc agggttttct 60tcaacgcatc agcctcacgg caaaacgcgc atgctgtaaa gttctcactt ccacccagc 119350175DNAGlycine max 350gaagaatcag agaatggcaa ctatgactgg tgtgagcctt tcatgcccca gggttttctt 60caacgcatca gcctcaccgc aaaacgcgca tgctgtaaag ttctcacttc cacccagcca 120agcagtgcga ccgggtagta tcaagttggg tcgcgtgatg aggatccgac ccgtt 175351285DNAGlycine max 351gaagaatcag agaatggcaa ctatgactgg tgtgagcctt tcatgcccca gggttttctt 60caacgcatca ggctcaccgc aaaacgcgca tgctgtaaag ttctctttta ttgtattttg 120tccagcagat tctatgactt gacaggactg cagaatggaa ttctgaagcg agggagagag 180attttcctca ctggttgcta cctccgaact cccactggag gttctggaca ttcacgtctt 240ttgccaacag agtatcttgt gattctattg gatgaagact tccaa 285352111DNAGlycine maxunsure at all n locations 352gaatggcaac tatgactggt gtgagccttt natgccccag ggttttcttc aacgcatnag 60cntcacnngn aaaacgcgca tgctgtaaag ttctcanttc cacacaacat a 111353156DNAGlycine max 353cttagacctc atcatcataa accccctcca gagaacaaga aacatccgaa tggcaactat 60gactggtgtg agcctttcaa gccccagggt tttcttcaac gcatcaccct caccgcaaaa 120cacgtacgcc gtaaagttcg cagttccact cagcca 156354136DNAGlycine max 354tcatcataaa ccccctccag agaacaagaa tcacagaatg gcaactatga ctggtgtgag 60cctttcaagc cccagggttt tcttcaacgc atcaccctca ccgcaaaaca cgtacgccgt 120aaagttcgca gttcca 13635585DNAGlycine maxunsure at all n locations 355ctatgactgg tgtgagcctt tcaagcccca gggttntctt caacgcatca ccctcacngc 60aaaacacgta cgccgtaaag ttcgc 85356356DNAGlycine max 356ctctctgaaa tgggtttcgc tttggcatac acagcatctg gttgttgctc aaacctacaa 60tttcagtctc tgttattcgc tgctgcttca ttgagatcaa aaccgtgtct ctctctctgc 120aactctactt atcgacccaa acgcattctc cagcgttctc caattgttgg cgctcagtct 180gaaaatggag ctctggttac ttcggagaag cccgacacta attacggaag acaatacttc 240cccctcgctg ctgttgtagg ccaagattct ataaaaactg ctcttttact tggtgcaatt 300gaccccgggg ttggaggaat tgccatatca ggaaagcgag gaactgccaa aactgt 356357339DNAGlycine maxunsure at all n locations 357anatgggttt cgctttggca ttcacagctt cttctacttg ctgntcaaat ctacaatctc 60agtctctgtt attcgctgct gctgcattga gatcaaaacc gtgtctctct ctctgcaaca 120cttatcgacc caaacgcatt cggaagcgtt ctcnaattgt tggcgctcaa tctgaaaacg 180gagctctcgt tacttccgag aagcctgaca ctaattacgg nagacaatac ttccccctcg 240ctgctgttgt aggccaagat gctataaaaa ctgctctttt acttggggcc attgaccctg 300ggattggagg aattgccata tcatgaaagc gaggnactg 339358284DNAGlycine maxunsure at all n locations 358tccggttatg gcgtccgcct tgggcacttc ttcaattgcn gttctgcctt cgcgctactt 60ctcttcttct tcctcccagc cttccattca cactctctct nnaacttcag ggcagaacta 120tgggcggaag ttttatggag gaattggaat ccatggcata aagggaaggg ctcagctctc 180ggttaccaat gttgccactg aagttaactc tgnagaacag gctcagagta ttgcttctaa 240aganagccag aggccagtat acccattttc tgccatantt ggnc 284359263DNAGlycine max 359tggcgtccgc cttgggcact tcttcaattg cggttctgcc ttcgcgctac ttctcttctt 60cttcttccaa gccttccatt cacactctct ctctaacttc agggcagaac tatgggcgga 120agttttatgg aggaattgga atccatggca taaagggaag ggctcagctc tcggttacca 180atgttgccac tgaagttaac tctgtagaac aggctcagag tattgcttct aaagaaagcc 240agaggccagt atacccattt tct 263360280DNAGlycine maxunsure at all n locations 360gtctgtcttc tcttctcttc tccggttatn gcgtccgcct tgggcacttc ttcaattgcg 60gttctgcctt cngggtactc tcttcttctt cttccaagcc ttccattcac actctctctc 120taacttcagg gcagaactat gggcggaagt tttatggagg aattggaatc catggcataa 180agggaagggc tcagctctcg gttaccaatg ttgccactga agttaactct gtagaacagg 240ctcagagtat tgcttctaaa gaaagccaga ggccagtata 280361278DNAGlycine maxunsure at all n locations 361tctgctccgg ttatggcntc cgncttgggc acttcttcaa ttgcngntct gccttncncg 60ctacttctct ncntcttctt ccaagccttc cattcanact cnctctctaa cttcanggca 120gaactatggg cggaagtttt atggaggaat tggaatccat ggnataaang gaagggctca 180gctctcggtt accaatgttg ncantgnagt taactctgna naacaggctc agantattgc 240ttctaaagaa agccagaggc cagtataccc attttctg 278362283DNAGlycine max 362attgctacat agcacacact ccctcttctc ttctacggtt atggcgtcca cgttgggcac 60ttcttcaatt gcggttcttc cttcgcgctg catctcttct ttttcttcca agccttccat 120tcacacactc tctctaactt cagggcagag ctatgggcgg aaattttatg gaggaattgg 180aattcatggc atcaagggaa ggtctcagct ctcagttgcc aatgttgcca ctgaagttaa 240ctctgtagaa caggcccaaa gtattgcttc taaagaaagc cag 283363273DNAGlycine maxunsure at all n locations 363gnaacaaatt gctacatagc acacactccc tcttctcttc tacggttatg gcgtccacgt 60tgggcacttc ttcaattgcg gttcttcctt cgcgctgcat ctcttctttt tcttccaagc 120cttccattca cacactctct ctaacttcag ggcagagcta tgggcggaaa ttttatgnag 180gaattggaat tcatggcatc aagggaaggt ctcagctctc agttgccaat gttgccactg 240aagttaactc tgtagaacag gcccaaagta ttg 273364259DNAGlycine max 364caaattgcta catagcacac actccctctt ctcttctacg gttatggcgt ccacgttggg 60cacttcttca attgcggttc ttccttcgcg ctgcatctct tctttttctt ccaagccttc 120cattcacaca ctctctctaa cttcagggca gagctatggg cggaaatttt atggaggaat 180tggaattcat ggcatcaagg gaaggtctca gctctcagtt gccaatgttg ccactgaagt 240taactctgta gaacaggcc 259365253DNAGlycine max 365acggctgcga aagacgacag aaggggacgg ttatggcgtc cacgttgggc acttcttcaa 60ttgcggttct tccttcgcgc tgcatctctt ctttttcttc caagccttcc attcacacac 120tctctctaac ttcagggcag agctatgggc ggaaatttta tggaggaatt ggaattcatg 180gcatcaaggg aaggtctcag ctctcagttg ccaatgttgc cactgaagtt aactctgtag 240aacaggccca aag 253366243DNAGlycine maxunsure at all n locations 366aataaaagac aaaagaaaca aaangctaca tagcatacag tctgtcttct cttctcttct 60ccggttatgg cgtccgcctt gggcacttct tcaattgcgg ttctgccttc gcgctacttc 120tcttcttctt cttccaagcc ttccattcac actctctctc taacttcagg gcagaactat 180gggcggaagt tttatggagg aattggaatc catggcataa agggaagggc tcagctctcg 240gtt 243367259DNAGlycine maxunsure at all n locations 367gcacacactc cctcttctct tctacggtta tggcgtccac gttgggcact tcttcaattg 60cggttcttcc ttcgcgctgc atctcttctt tttcttccaa gccttccatt cacacactct 120ctctaacttc agggcagagc tatgggcgga aattttatgg aggaattgga attcatgggc 180atcaagggaa ngtctcagct ctcagttgcc aatgttgcca ctgaagttaa ctctgtagaa 240caggcccaaa gtattgctt 259368163DNAGlycine max 368caaattgcta catagcacac actccctctt ctcttctacg gttatggcgt ccacgttggg 60cacttcttca attgcggttc ttccttcgcg ctgcatctct tctttttctt ccaagccttc 120cattcacaca ctctctctaa cttcagggca gagctatggg cgg 163369151DNAGlycine max 369gaaattgcta catagcacac actccctctt ctcttctacg gttatggcgt ccacgttggg 60cacttcttca attgcggttc ttccttcgcg ctgcatctct tctttttctt ccaagccttc 120cattcacaca ctctctctaa cttcagggca g 151370232DNAGlycine max 370gaagaatgaa atctatctat cttcttatcc gaagcccgtg aggccaataa gaagcacgtc 60agctgctatg aatggtgaat aaaacacaaa agaaacaaat tgctacatag cacacactcc 120ctcttctctt ctacggttat ggcgtccacg ttgggcactt cttcaattgc ggttcttcct 180tcgcgctgca tctcttcttt ttcttccaag ccttccattc acacactctc tc 232371107DNAGlycine max 371tacggctgga agacgacaga agggggaata aaacacaaaa gacacaaatt gctacatagc 60acacactccc tcttctcttc tacggttatg gcgtccacgt tgggcac 107372235DNAGlycine max 372ctcgagccga atcggctcga ggcagattaa aagggatgga attaccaagc ttgttattct 60tccactttat ccacaatttt caatatcaac cagtggctca agcctacgtc tactggagag 120tatattccga gaggatgagt atctagtcaa catgcagcac acagtaatac catcatggta 180tcaacgtgaa ggatacataa aggccatggc aaatttgatt gagaaagagt tgaga 235373250DNAGlycine max 373gaccaggcac ttgcaattaa aatggctttg gaagcaaagg gcatctcttc aaatgtctac 60gttgggatgc gatactggta cccatttacc gaagaagcaa ttcagcaaat taagagggac 120agaataacaa ggcttgtggt actacccctt tatccccagt tttctatatc cacaactgga 180tcaagcatcc gtgttcttga gcatatattc agggaagatg cctacttgtc taagctccct 240gtttccatta 250374254DNAGlycine max 374ggaatgtgtt gatttgatca tggaagagct tgaaaagaga aagataacta atgcatacac 60ccttgcttat cagagtagag ttggacctgt ggaatggtta aaaccctata cagatgagac 120aataattgaa cttgggaaaa agggagtaaa aagcctgctg gctgtaccaa ttagctttgt 180cagcgagcat attgaaactc tcgaagaaat tgatgttgag tacaaagaat tggctctaaa 240ctctggtata gaaa 254375248DNAGlycine max 375gaaaaagttg gtgtgctgct tctcaatcta ggaggaccag agacattgaa tgacgttcaa 60ccttttctgt ttaatctttt tgcagatcct gatatcattc gtcttccaag gttgtttcgg 120tttctccagc gaccattggc aaaattgatt tctgtacttc ggtctcctaa atccaaggaa 180gggtatgctg ctattggtgg tggctctcct ttacgcaaaa ttacagatga ccaggcactc 240gcaattaa 248376275DNAGlycine max 376aattgacatg gagtacaagg aattggctct tgaatctggc atcaagaatt gggcacgtgt 60acctgccctt ggtgttaccc cttccttcat tacagattta gcagatgcag taatagaagc 120tctcccatca gcaacagcaa tatatgcacc gaccagaacc tctgaagatg ttgatcatga 180cccagttaga tattttatca agatgttctt tggttcaatc ttggcattca tcttgttctt 240gtcacccaaa atgatcacgg cattcaggaa tcatg 275377288DNAGlycine max 377ccttccttca tacagattta gcagatgcag taatagaagc tctcccatca gcaacagcaa 60tatatgcacc gaccagaacc tctgaagatg ttgatcatga cccagttaga tattttatca 120agatgttctt tggttcaatc ttggcattca tcttgttctt gtcacccaaa atgatcacgg 180cattcaggaa tcatgtcatt tagaagaatt aaatcctgct tgctgaattc aatctgcaag 240catatagatg aagcctattg atagcaacaa agtatacttt gatttttt 288378282DNAGlycine max 378atggaaaaaa gggagtgaaa agtctgctcg ctgttccaat tagcttcgtc agtgagcata 60ttgaaactct agaagaaatt gatgttgaat acaaagagtt ggctctagaa tctggtatag 120aaaagtgggg ccgtgttcct gctctaggat gcgaacctac cttcatttct gatttggcag 180atgccgttat tgagagtctc ccatatgttg gtgccatgac agcttcagac cttgaagctc 240aacaatcctc gttccatggg cagtgtagaa gagttattgg ca 282379237DNAGlycine max 379catccgtgtt cttgagcata tattcaggga agatgcctac ttgtctaagc tccctgtttc 60cattataaac tcttggtatc aacgagaagg ttatattaag tcaatggcta acttaattca 120gaaagagctc cagagttttt ctgaaccaaa agaggtaatg atatttttca gtgcccatgg 180tgtacctgtc agttacgttg aggaagctgg ggatccatac cgagaccaaa tggagga 237380253DNAGlycine max 380actggatcaa gcatccgtgt tcttgagcat atattcaggg aagatgccta cttgtctaac 60ctccctgttt ccattataaa ctcttggtat caacgagaag gttatattaa gtcaatggct 120aacttaattc agaaagagcg ccagagtttt tcttaaccaa aagaggtaat gatatttttc 180agtgcccatg gtgtacctgt caagtacgtt gagggagctg gggatccata ccgagaccaa

240atggaggagt gca 253381269DNAGlycine max 381ttcttgagca tatattcagg gaagatgcct acttgtctaa gctccctgtt tccattataa 60actcttggta tcaacgagaa ggttatatta agtcaatggc taacttaatt cagaaagagc 120tccagagttt ttctgaacca aaagaggtaa tgatattttt cagtgcccat ggtgtacctg 180tcagttacgt tgaggaagct ggggatccat accgagacca aatggaggag tgcatcttct 240tgatcatgca agagttgaaa gctagagga 269382251DNAGlycine max 382aagagctcca gagtttttct gaaccaaaag aggtaatgat atttttcagt gcccatggtg 60tacctgtcag ttacgttgag gaagctgggg atccataccg agaccaaatg gaggagtgca 120tcttcttgat catgcaagag ttgaaagcta gaggaattag taatgagcac actcttgctt 180atcagagtcg agtgggtcct gtacagtggc tgaaaccata tactgatgaa gttctcgttg 240agcttggcca a 251383275DNAGlycine max 383ttaattcaga aagagctcca gagtttttct gaaccaaaag aggtaatgat atttttcagt 60gcccatggtg tacctgtcag ttacgttgag gaagctgggg atccataccg agaccaaatg 120gaggagtgca tcttcttgat catgcaagag ttgaaagcta gaggaattag taatgagcac 180actcttgctt atcagagtcg agtgggtcct gtacagtggc tgaaaccata tactgatgaa 240gttctcgttg agcttggcca aaaaggtgtg aagag 275384168DNAZea mays 384ctttcttaca tatattcagc accacctctc aagctcgagc agaatggatg gattgggaac 60ttcgctctgg gtgcgagtta catcagcttg ccctggtggg ctggccaggc gttatttgga 120actcttacac cagatatcag tgtcttgact actttgtaca gcatagct 168385256DNAZea mays 385attgaagggg ataggactct ggggcttcag tcacttcctg ttgcttttgg gatggaaact 60gcaaaatgga tttgtgttgg agcaattgat atcactcaat tatctgttgc aggttaccta 120ttgagcaccg gtaagctgta ttatgccctg gtgttgcttg ggctaacaat tcctcaggtg 180ttctttcagt tccagtactt cctgaaggac cctgtgaagt atgatgtcaa atatcaggca 240agcgcacaac cattct 256386411DNAZea mays 386cccacgcgtc cgcccacgcg tccgcccacg cgtccgccca cgcgtccgag cacacacggg 60cgcatcaggg cctagctcga gtccactact tgaaaaacag gaaaaaggtt gcgtttgagg 120agatgacgaa gctcgtggag atagccagcc actgcgcgtc ggcatatgaa aagcggtcgg 180aatacggtga gcgcgaagct gcgaggagcg acctgaacat ggcgacgctt cttgatccta 240ccaggactta tccttacaga tacagagcag ctgtactgat ggacgaaggc aaggaggagg 300aggcgatcgc ggagctgtca ggagccatag ctttcaagcc ggaccttcag ctgctgcacc 360ttcgcgcggc gttcttcgac tccatgggcg agcgcgagag cgccctgtgg g 411387484DNAZea maysunsure at all n locations 387ntggggttnn ctagagggga ggggggcaat tgatggaagt cttcaattcc gtttcgnacc 60nncccgcccc acgcgtccgc cgacgccaaa aacgcgaagg cgaacgccat ggccccgaat 120aagagcaccc gcggcggatg actccagttt caaccagctg ctcggtatca aaagtgctta 180gccagggaac ggccttttgg aaaatccgcc ttaacttaac taagccggtg acatggcctc 240cgcttgtttg gggagttctc tgtggagcag ctgcctctgg aaatttccac tggacagttg 300aagatgtcgc aaaatctatt gtatgcatga taatgtctgg tccatgcctt acaggataca 360cacagacact taatgactgg tatgatcgag acattgatgc aattaatgag ccttatcggc 420ctattccatc aggtgctata tcaganaacg aggtaataac ccagatctgg gtgctattgc 480tagg 484388301DNAZea mays 388ccaaggcccc gaataacgca cccgcggcgg atggctccag tttcaaccag ctgctcggta 60tcaagggtgc taagcaagac agcgacatgt ggcagatgcg tcttcaactt actaagccgg 120tgacatggcc tccgcttgtt tggggagttc tctgtggagc agctgcctct ggaaatttcc 180agtggacagt tgaagatgtc gcaaaatcta ttgtatgcat gataatgtct ggtccatgcc 240ttacaggata cgcacagaca cttaatgact ggtatgatcg agacattgat gcaattagtg 300a 301389284DNAZea mays 389tgaagatgtc gcaaaatcta ttgtatgcat gataatgtct ggtccatgcc ttacaggata 60cacacagaca cttaatgact ggtatgatcg agacattgat gcaattaatg agccttatcg 120gcctattcca tcaggtgcta tatcagaaaa cgaggtaata acccagatct gggtgctatt 180gctaggaggg cttggattgg gtgctttgtt agatgtgtgg gcaggacatg attttcctat 240tgtgttttat cttgctgtgg gtggctcctt actttcttac atat 284390256DNAZea mays 390caattaatga gccttatcgg cctattccat caggtgctat atcagaaaac gaggtaataa 60cccagatctg ggtgctattg ctaggagggc ttggattggg tgctttgtta gatgtgtggg 120caggacatga ttttcctatt gtgttttatc ttgctgtggg tggctcccta ctttcctaca 180tatattcagc accacctctc aagctccagc agaatggatg gaatgggaac ttcgctctgg 240gtgcgagtta catcag 256391318DNAZea mays 391gcatgataat gtctggtcca tgccttacag gatacacaca gacacttaat gactggtatg 60atcgagacat tgatgcaatt aatgagcctt atcggcctat tccatcaggt gctatatcag 120aaaacgaggt aataacccag atctgggtgc tattgctagg agggcttgga ttgggtgctt 180tgttagatgt gtgggcagga catgattttc ctattgtgtt ttatcttgct gtgggtggct 240ccttactttc ttacatatat tcagcaccac ctctcaagct caagcagaat ggatggattg 300ggaacttcgc tctgggtg 318392272DNAZea mays 392ctggtgtaag agttccaaat aacgcctggc cagcccacca gggcaagatg atgtaactct 60aacccagagc gaagttccca atccatccat tctgcttgag cttgagaggt ggtgctgaat 120atatgtaaga aagtaaggag ccacccacag caagataaaa cacaatagga aaatcatgtc 180ctgcccacac atctaacaaa gcacccaatc caagccctcc tagcaatagc acccagatct 240gggttattac ctcgttttct gatatagcac ct 272393288DNAZea mays 393cacacagaca cttaatgact ggtatgatcg agacattgat gcaattaatg agccttatcg 60gcctattcca tcaggtgcta tatcagaaaa cgaggtaata acccagatct gggtgctatt 120gctaggaggg cttggattgg gtgctttgtt agatgtgtgg gcaggacatg attttcctat 180tgtgttttat cttgctgtgg gtggctcctt actttcttac atatattcag caccacctct 240caagctcaag cagaatggat ggattgggaa cttcgctctg ggtgcgag 288394256DNAZea mays 394caattcctca ggtgttcttt cagttccagt acttcctgaa ggaccctgtg aagtatgatg 60tcaaatatca ggcaagcgca caaccattct tcgtactggg cctactggtg acagcactgg 120caaccagcca ttaatgaagg caaacttaaa cagaacgagc aaccgttctg ataccgaaga 180ggcacgtctg gtgaccatta ataagctagc tgcttgtgtg cagggtagga agagaacgtc 240tttttacttg tagaac 256395280DNAZea mays 395caattcctca ggtgttcttt cagttccagt acttcctgaa ggaccctgtg aagtatgatg 60tcaaatatca ggcaagcgca caaccattct tcgtactggg cctactggtg acagcactgg 120caaccagcca ttaatgaagg caaacttaaa cagaacgagc aaccgttctg ataccgaaga 180ggcacgtctg gtgaccatta ataagctagc tgcttgtgtg cagggtagga agagaacgtc 240tttttacttg tagaacacag atcgattttg taagggttat 280396287DNAZea mays 396cccacgcgtc cgtattcagc accacctctc aagctcaagc agaatggatg gattgggaac 60ttcgctctgg gtgcgagtta catcagcttg ccctggtggg ctggccaggc gttatttgga 120actcttacac cagatatcat tgtcttgact actttgtaca gcatagctgg gctagggatt 180gctattgtaa atgatttcaa gagtattgaa ggggatagga ctctggggct tcagtcactt 240cctgttgctt ttgggatgga aactgcaaaa tggatttgtg ttggagc 287397152DNAZea mays 397cagcaccacc tctcaagctc aagcagaatg gatggattgg gaacttcgct ctgagtgcga 60gttacatcag cttgccctgg tgggctggcc aggcgttatt tggaactctt acaccagata 120tcattgtcta gactacttcg tacagcatag ct 152398298DNAZea mays 398agggcttcgt gtcggaggcg gagtccggca agaggctggc gcaggtggtc agcgacccca 60gcctcaccaa gtcgggggtg tactggagct ggaacaagga ctcggcgtcg ttcgagaacc 120agctgtcgca ggaggccagc gatccggaga aggccaagaa gctctgggag atcagcgaga 180agctcgtggg gcttccttga gctccccgca caggaaaaag cgacatgatg aatctgtcga 240gcagaggagc tttcgcttcg ttgtattatg tgtaacatta gcatccattt gtttgttt 298399218DNAZea mays 399ggggagttcg acggcgccaa ggcatacaag gacagcaagg tgtgcaacat gctgacgatg 60caggagttcc accgccggta ccacgaggag acgggcgtga ccttcgcgtc gctctacccg 120ggctgcatcg ccaccagggg cctgttccgc gaacaaattc cgctgttccg gctgtgctcc 180gcccgccgtt ccagaagtac atcaccaggg tacgtctc 218400317DNAZea mays 400gtcacttctc cacgaacaaa agcgcatcga tctcgctgtc gtcactcctc gtcacccagc 60cacgaacaga ggcaccaccc agcatggccc tgcaggcggc gctactccca tacaccctct 120catccgtccc caagaagtgc agcctcgccg tcgcggcgaa tgacacggca ttccttagcg 180tatcctacaa gaaggtgcac gcggcgtcac tgtccgtgaa aacgcggtgg cgactaccgc 240gcctgtggcc acgccggggt ccagcacggc ggtcaacgat gggaagaaga ccgtgcggca 300tgccgtggtg gtgatca 317401172DNAZea mays 401gcagaagtcc gactacccgt cccggcggct tatcatcctc gggtccatca ccggcaacag 60caacacgctg gccgggaaca tcccgcccaa ggccgggctg ggcgaccttc gcgggctcgc 120ggcggggctg cgcggccaga acggctctgc catgatcgac ggcttcgaga gc 172402313DNAZea mays 402aaatcctcag tcctcaggct gctcacagtt cgtgctatcc gctcgcgctc ccggtagtct 60gcctgctcgg caattcggca tggcgctcca ggccgcgacg tccttcctcc cctcggccct 120ctcggcgcgc aaggaggggt cgtcggtgaa ggactcggcg ttcttgggtg tccatctcgc 180ggacgatggc ctcaagctgg agaccgctgc tctgggccta cgcaccaaga gggtgatcac 240gtcggtggcc atccgcgcgc aggcggcagc ggtgtcctca ccatcagtat accccgcgtc 300gccgtccggc aag 313403252DNAZea mays 403cccagccaaa tcctcagtcc tcaggctgct cacagttcgt gctatccgct cgcgctcccg 60gtagtctgcc tgctcggcaa ttcggcatgg cgctccaggc cgcgacgtcc ttcctcccct 120caggccctct gcggcgcgca aggtaggggt cgtcggtgaa ggactcggcg ttcttgggtg 180tccatctcgc ggacgatggc ctcaagctgg agaccgctgc tatgggccta cgcaccaaga 240gggtgatcac gt 252404399DNAZea mays 404accacgcgtc cgcatacaag gacagcaagg tgtgcaacat gctgacgatg caggagttcc 60accgccggta ccacgaggag acgggcgtga ccttcgcgtc gctctacccg ggctgcatcg 120ccaccacggg cctgttccgc gagcacatcc cgctgttccg cctgctcttc ccgccgttcc 180agaagtacat caccaagggg tacgtctccg aggaggaggc cgggaagcgg ctggcgcagg 240tggtgagcga ccccagcctg accaagtccg gcgtgtactg gagctggaac aagaactccg 300cgtccttcga gaaccagctc tctgaggagg ccagcgacgc cgacaaggcc aagaagctct 360gggagatcag cgagaagctc gtcggcttgg cgtgatgcc 399405442DNAZea maysunsure at all n locations 405acaccggcac accaacacgc tggccgggaa catcccgccc aaggccgggc tgggcgacct 60ccgcggcgtg gcggcggngc tgcgcggcca gaacggctct gccatgatcg acggctccga 120gagcttcgac ggcgccaagg cgtacaagga cagcaagatc tgcaacatgc taacaatgca 180ggagctgcac cggcggtacc acgaggagac gggcatcacg ttcgcgtcgc tctacccggg 240gtgcatcgcc accacggggc tgttccgcga gcacatcccg ctgttccggc tgctcttccc 300gccgttccag aagttcgtca ccaaaggctt cgtgtcggaa gcggagtccg gcaagaagct 360ggcgcatgtg gtcagcgacc ccagcctcac caagtcggng gtgtactgga gctggaacaa 420ggactccgcg tcgttcgaga ac 442406442DNAZea mays 406gcgatcacgg gcgacgccaa cacgctggcc ggtgacatct cgcccaaggc cgggctgggc 60gacctccgcg gcctcgcggc ggggctgcgc ggccagaacg gctctgccat gatcgacggc 120tccgagagct tcgacggcgc caaggcgtac aaggacagca agatctgcaa catgctcacc 180atgcaggagc tgcaccggcg gtaccacgag gagacgggca tcacgttcgc gtcgctctac 240ccggggtgca tcgccaccac ggggctgttc cgcgagcaca tcccgctgtt ccgcctgctc 300ttcccgcctt tccagaagtt cgtcaccaag ggcttcgtgt cggaggcgga gtccggcaag 360aggctggcgc atgtggtcag cgaccccagc cttaccaaag tcggggtgta ctggagctgg 420aacaggggac tcgcgtcgtt cg 442407352DNAZea mays 407ctcctggcgc gcctgctcct ggacgacatg cagaagtccg actacccgtc ccggcgagtc 60atcatcctcg gctccatcac cggcaacacc aacacgctgg ccgggaacat cccgcccaag 120gccgggctgg gcgacctgcg cggcctcgcg gcggggctgc gcggccagaa cggctctgcc 180atgatcgacg gctccgagag cttcgacggc gccaaggcgt acaaggacag caagatctgc 240aacatgctca ccatgcagga gctgcaccgg cggtaccacg aggagacggg catcacgttc 300gcgtcgctct acccggggtg catcgccacc acggcgctgt tccgcgagca ca 352408277DNAZea mays 408ctggccggga acatcccgcc caaggccggg ctgggcgacc tccgcggcct cgcggcgggg 60ctgcgcggcc agaacggctc tgccatgatc gacggctccg agagcttcga cggcgccaag 120gcgtacaagg acagcaagat ctgcaacatg ctaacaatgc aggagctgca ccggcggtac 180cacgaggaga cgggcatcac gttcgcgtcg ctctacccgg ggtgcatcgc caccacgggg 240ctgttccgcg agcacatccc gctgttccgg ctgctct 277409272DNAZea mays 409gacggcgcca aggcatacaa ggacagcaag gtgtgcaaca tgctgacgat gcaggagttc 60caccgccggt accacgagga gacgggcgtg accttcgcgt cgctctaccc gggctgcatc 120gccaccacgg gcctgttccg cgagcacatc ccgctgttcc gcctgctctt cccgccgttc 180cagaagtaca tcaccaaggg gtacgtctcc gaggaggagg ccgggaagcg gctggcgcag 240gtggtgagcg accccagcct gaccaagtcc gg 272410309DNAZea mays 410cactggccgg gaacatcccg cccaaggccg ggctgggcga cctccgcagc ctcgcggcgg 60ggctgcgcgg ccagaacggc tctgccatga tcgacggctc cgagagcttc gacggcgcca 120aggcgtacaa ggacagcaag atctgcaaca tgctcaccat gcaggagctg caccggcggt 180accacgagga gacgggcatc acgttcgcgt cgctctaccc ggggtgcatc gccaccacgg 240ggctgttccg cgagcacatc ccgctgttcc gcctgctctt cccgccgttc cagaagttcg 300tcaccaagg 309411264DNAZea mays 411cagaacggct ctgccatgat cgacggctcc gagagcttcg acggcgccaa ggcgtacaag 60gacagcaaga tctgcaacat gctcaccatg caggagctgc accggcggta ccacgaggag 120acgggcatca cgttcgcgtc gctctacccg gggtgcatcg ccaccacggg gctgttccgc 180gagcacatcc cgctgttccg cctgctcttc ccgcctttcc agaagttcgt caccaagggc 240ttcgtgtcgg aggcggagtc cggc 264412267DNAZea mays 412gctcggtgat gatcgacggc ggggagttcg acggcgccaa ggcatacaag gacagcaagg 60tgtgcaacat gctgacgatg caggagttcc accgccggta ccacgaggag acggccgtga 120ccttcgggtc gctctacccg ggctgaatgg caacaacggg cctgttccgg gaacacatcc 180cgctgttccg gctgctcttc ccgccgttcc agaagtacat caccaagggg gtacgtctcc 240gaggaggagg ccgggaagcg ctggcgc 267413302DNAZea mays 413ggcgtacaag gacagcaaga tctgcaacat gctcaccatg caggagctgc accggcggta 60ccacgaggag acgggcatca cgttcgcgtc gctctacccg gggtgcatcg ccaccacggg 120gctgttccgc gagcacatcc cgctgttccg cctgctcttc ccgccgttcc agaagttcgt 180caccaagggc ttcgttccga agcggaaccg gcaagaagct tgcgcaggtg gtcagcgacc 240ccagcctcac caagtcgggg gtgtactgga gctggaacaa ggactcggcg tcgttcgaga 300ac 302414291DNAZea mays 414ggcgcgcctg ctcctggacg acatgcagaa gtccgactac ccgtcccgcc gcctcatcat 60cctcggctcc atcaccggca acaccaacac gctggccggg aacatcccgc ccaaggccgg 120gctgggcgac ctccgcagcc tcgggcgggg ctgcgcggcc agaacggctc tgccatgatc 180gacggctccg agagcttcga cggcgccaag gcgtacaagg acagcaagat ctgcaacatg 240ctaacaatgc aggagctgca ccggcggtac cacgaggaga cgggcatcac g 291415268DNAZea mays 415cgagcacatc ccgctgttcc gcctgctctt cccgccgttc cagaagtaca tcaccaaggg 60gtacgtctcc gaggaggagg ccgggaagcg gctggcgcag gtggtgagcg accccagcct 120gaccaagtcc ggcgtgtact ggagctggaa caagaactcc gcgtccttcg agaaccagct 180ctctgaggag gccagctgac gcgacaaggc caagaagctc tgggagatcc gcgagaagct 240cgtcggcttg gcgtgatgcc caccgtgc 268416296DNAZea mays 416cccacgcgtc cgaacacgct ggccgggaac atcccgccca aggccgggct gggcgacctc 60cgcggcctcg ggcggggctg cgcggccaga acggctctgc caggatcgac ggctccgaga 120gcttcgacgg cgccaaggcg tacaaggaca gcaagatctg caacatgctc accatgcagg 180agctgcaccg gcggtaccac gaggagacgg gcatcacgtt cgcgtcgctc tacccggggt 240gcatcgccac cacggggctg ttccgcgagc acatcccgct gttccgcctg ctcttc 296417255DNAZea mays 417gcctgctctt cccgccattc cagaagtaca tcaccaaggg gtacgtctcc gaggaggagg 60ccgggaagcg gctgtcgcag gtcgtgagcg accccagcct gaccaagtcc ggcgtgtact 120ggagctggaa caagaactcg gcgtccttcg agaaccagct ctctgaggag gccagcgacg 180ccgacaaggc caagaagctc tgggagatca gcgagaagct cgtcagcttg gcgtgacgac 240ctgatgtcca cagtg 255418326DNAZea mays 418cggacgcgtg ggcggacgcg tggggaagta catcaccaag gggtacgtct ccgaggagga 60ggccgggaag cggctggcgc aggtggtgag cgaccccagc ctgaccaagt ccggcgtgta 120ctggagctgg aacaagaact ccgcgtcctt cgagaaccag ctctctgagg aggccagcga 180cgccgacaag gccaagaagc tctgggagat cagcgagaag ctcgtcggct tggcgtgatg 240cccaccgtgg ccggcgccgg cagccggcga cagtttttcc tacctaggac atgctcatta 300gttggtctca gtcgagtagt cgacgt 326419290DNAZea mays 419ctccgaggag gaggccggga agcggctgtc gcaggtcgtg agcgacccca gcaccgacca 60agtccggcgt gtactggagc tggaacaaga actcggcgtc cttcgagaac cagctctctg 120aggaggccag cgacgccgac aaggccaaga agctctggga gatcagcgag aagctcgtcg 180gcttggcgtg acgacctgat gcccaccgtg gccggcgccg gcagccggtg acagtttttt 240cctaggacat gttcgttact tgatctcagt cgacgcgtgg tgcactcgtg 290420217DNAZea mays 420cccacgcgtc cgctgggcca cttcctcctg gcgcgcctgc tcctggacga catgcagaag 60tccgactacc cgtcccgccg cctcgtcatc ctcggctcca tcaccggcaa caccaacacg 120ctggccggga acatcccgcc caaggccggg ctgggcgacc tccgcggcct cgcggcgggg 180ctgcgcggcc agaacggctc tgccatgatc gacggct 217421242DNAZea mays 421ctccgaggag gaggggaagc ggctggcgca ggtggtgagc gaccccagcc tgaccaagtc 60cggcgtgtac tggagctgga acaagaactc cgcgtcctac gagaaccagc tctctgagga 120ggccagcgac gccgacaagg ccaagaagct ctgggagatc agcgagaagc tcgtcggctt 180ggcgtgatgc ccaccgtggc cggcgccggc agccggcgac agtttttcct acctaggaca 240tg 242422116DNAZea mays 422tgccggtacc acgaggagac gggcgtgacc ttcgcgtcgc tctacccggg ctgcatcgcc 60accacgggcc tgttccgcga gcacatcccg ctgttccgcc tgctcttccc gccgtt 116423133DNAZea mays 423tctcgagccg aatctggctc gaggaggaac atcccgccca aggccgacct gggcgacctc 60cgacgcctcg cggcggggct gcacggccat aacggctctg ccatgatcga cggctccgag 120agcttcgacg gcg 133424364DNAZea mays 424cgcaagggca cggcggtcat caccggcgcg tcgtccggcc tcggcctcgc cacggcgaag 60gccctggcgg agacaggcaa gtggcacgtc atcatggcct gccgcgactt cctcaaggcg 120tcgcgcgcgg ccaaggcggc cggcatggac aaggacagct tcaccgtcgt gcacctggac 180ctcgcctccc tggacagcgt ccgccagttc gtcaagaacg tgcgccagct ggagatgccc 240atcgacgtgg tggtctgcaa cgccgtcgtg taccagccca ccgccaagga gccgtcctac 300accgccgacg gcttcgagat gagcgtcggc gtcaaccaac ctggccactt tctcctcgcg 360cgcg 364425289DNAZea mays 425cctggacctc gcctccctgg acagcgtccg ccagttcgtc aggaacgtgc gccactgaga 60gatgcccatc gacgtggtgg tctgcaacgc cgccgtgtac cagcccaccg ccaaggagcc 120gtcctacacc gccgacggct tcgagatgag cgtcggcgtc aaccacctcg gccacttcct 180cctcgcgcgc gagctcctca

gcgacctcca gtcctccgac tacccctcta agcgcctcat 240catcgtcggc tccatcaccg ggaacacgta cacgctggcg gggaacgtg 289426331DNAZea mays 426atccgcacac gcgtccgcgt catcatgggc tgccgcgatt tccacaaggc gtcgcgcgca 60gccaaagcag ccggcatgga caaggacagc ttcaccgtcg tgcacctgga cctcgcctcc 120ctcgacagcg tccgccagtt cgtcaagaac gtgcgccagc tggagatgcc cgtcgacgtg 180gtggtctgca acgccgccgt gtaccagccc accgccaagg agccgtccta caccgccgac 240ggcttcgaga tgagcgtcgg cgtcaaacac ctcggccact tcctcctcgc ccgcgagctc 300ctcagcgacc tccagtcctc cgactatccc t 331427280DNAZea mays 427gtggtggtct gcaacgccgc cgtgtaccag cccaccgcca aggagccgtc ctacaccgcc 60gacggcttcg agatgagcgt cggcgtcaac cacctcggcc atttcctcct cgcccgcgag 120ctcctcagcg acctccagtc ctccgactac ccctctaagc gcctcatcat cgtcggctcc 180atcaccggga acacgaacac gctggcgggg aacgtgcccc cgaactcgaa cctgggcgac 240ctgcgcggcc tcgccggcgg cctcaacggc gttggcagct 280428285DNAZea mays 428gagcgtcggc gtcaaccacc tcggccattt cctcctcgcc cgcgagctcc tcagcgacct 60ccagtcctcc gactacccct ctaagcgcct catcatcgtc ggctccatca ccgggaacac 120gaacacgctg gcggggaacg tgcccccgaa ggcgaacctg ggcgacctgc gcggcctcgc 180cggcggcctc aacggcgttg gcagctcggt gatgatcgac ggcggggagt tcgacggcgc 240caaggcatac aaggacagca aggtgtgcaa catgctgacg atgca 285429282DNAZea mays 429cccacgcgtc cgcaccggcg cgtcgtccgg cctcggcctc gccacggcga aggccctcgc 60ggagacaggc aagtggcacg tcatcatggc ctgccgcgac ttcctcaagg cgtcgcgcgc 120ggccaaggcg gccggcatgg acaaggacag cttcaccgtc gtgcacctgg acctcgcctc 180cctggacagc gtccgccagt tcgtcaggaa cgtgcgccag ctggagatgc ccatcgacgt 240ggtggtctgc aacgccgccg tgtaccagcc caccgccaag ga 282430276DNAZea mays 430cccacgcgtc cggtcaggaa cgtgcgccac tggagatgcc catcgacgtg gtggtctgca 60acgccgccgt gtaccagccc accgccaagg agccgtccta caccgccgac ggcttcgaga 120tgagcgtcgg cgtcaaccac ctcggccatt tcctcctcgc ccgcgagctc ctcagcgacc 180tccagtcctc cgactacccc tctaagcgcc tcatcatcgt cggctccatc accgggaaca 240cgaacacgct ggcggggaac gtgccccgac agcgaa 276431229DNAZea mays 431ccaaaacctg cagagggtga gcaggtcggc ggacatccgc gcgcagacgg cagcggtgtc 60ctccccgtca gtgacccccg cgtcgccgtc tggcaagaag accctccgca agggcacggc 120ggtcatcacc ggcgcgtcgt ccggcctcgg cctcgccacg gcgaaggccc tcgcggagac 180aggcaagtgg cacgtcatca tggcctgccg cgacttctca aggcgtcgc 229432394DNAZea mays 432aggaagaacc cagccaaatc ctcagtcctc aggctgctcg cagctcgtgc cgtccactct 60cccccgaggc attctcttgc gttcgctgct cgacatggcg ctccaggcgg cgacgtcctt 120cctcccctct gccctctccg cgcgcaagga ggggtcggtg aaggactcgg cgtcgttctt 180gggtgttcgt ctcgcggcgg atgggctcaa gctggacacc accgctctgg gcctacgcac 240cgtgagggtg agcaggtcgg cggacatccg cgcgcagacg gcagcggtgt cctccccgtc 300agtgacccct gcgtcgccgt ctggcaagaa gaccctccgc attggcacgg cggtcatcat 360cggcgcgtcg tccggcctcg gcctcgccac ggcg 394433275DNAZea mays 433gttcgtctcg cggcggatgg cctcaagctg gacaccaccg ctctgggcct acgcaccgtg 60agggtgagca ggtcggcgga catccgcgcg cagacggcag cggtgtcctc cccgtcagtg 120acccccgcgt cgccgtctgg caagaagacc ctccgcaagg gcacggcggt catcaccggc 180gcgtcgtccg gcctcggcct cgccacggcg aaggccctcg cggagacagg caagtggcac 240gtcatcatgg cctgccgcga cttcctcaag gcgtc 275434418DNAZea maysunsure at all n locations 434agaggaagaa gaagaaccca gccaaatcct cagtcttcag gctgctcaca gctcgtgccg 60tccactctcc cccgaggcag tctcttgcgt tcgctgctcg acatggcgct ccaggcggcg 120acgtcctttc tcccctcggc cctctccgcg cgcaaggagg ggtcggtgaa ggactcggcg 180tcgttcttgg gtgttcgtct cgcggcggat ggcctcaagc tggacaccac cgctctgggc 240ctacgcaccg tgagggtgag caggtcggcg gacatccgcg cgcagacggc agcggtgtcc 300tcnccgtcag tgacncccgc gtccccgtct ggcaanaaga cctccgnaag ggnaanggcg 360gtcatnaacg gggggctngn tagggcncng gggnnncnna gggngaaggg ngccncnt 418435321DNAZea mays 435agccaaatcc tcagtcttca ggctgctcac agctcgtgcc gtccactctc ccccgaggca 60gtctcttgcg ttcgctgctc gacatggcgc tccaggcggc gacgtccttt ctcccctcgg 120ccctctccgc gcgcaaggag gggtcggtga aggactcggc gtcgttcttg ggtgttcgtc 180tcgcggcgga tggcctcaag ctggacacca ccgctctggg cctacgcacc gtgagggtga 240gcaggtcggc ggacatccgc gcgcagacgg cagcggtgtc ctccccgtca gtgaccccgc 300gatcgcgtct ggcaagaaga c 321436112DNAZea mays 436ctcgcccgcg agctcctcag cgacctccag tcctccgact actcctctaa gcgcctcatc 60atcgtcagct ccatcaccgg gaacacgaac acgctggcgg ggaacgtgcc cc 112437296DNAZea mays 437gactagttct agatcccccc gcggagcaga gaggaagaag aagaacccag ccaaatcctc 60agtcttcagg ctgctcacag ctcgtgccgt ccactctccc ccgaggcagt ctcttgcgtt 120cgctgctcga catggcgctc caggcggcga cgtcctttct cccctcggcc ctctccgcgc 180gcaaggaggg gtcggtgaag gactcggcgt cgttcttggg tgttcgtctc gcggcggatg 240gcctcaagct ggacaccacc gctctgggcc tacgcaccgt gagggtgagc aggtcg 296438175DNAZea mays 438cgacatggcg ctccaggcgg cgacgtcctt tctcccctcg gccctctccg cgcgcaagga 60ggggtcggtg aaggactcgg cgtcgttctt gggtgttcgt ctcgcggcgg atggcctcaa 120gctggacacc accgctctgg gcctacgcac cgtggaggtg agcaggtcag cggac 175439301DNAZea mays 439agaagaaccc agccaaatcc tcagtcctca ggctgctcac agctcgtgcc gtccactctc 60ccccgagcca gtctcttgcg ttcgctgctc gacatggcgc tccaggcggc gacgtccttc 120ctcccctctg ccctctccgc gcgcaaggag gggtcggtga aggactcggc gtcgttcttg 180ggtgttcgtc tcgcggcgga tggcctcaag ctggacacca ccgctctggg cctacgcacc 240gtgagggtga gcaggtcggc ggacatccgc gcgcagacgg cagcggtgtc ctccccgtca 300g 301440261DNAZea mays 440gtgaaggact cggcgtcgtt cttgggtgtt cgtctcgcgg cggatggcct caagctggac 60accaccgctc tgggcctacg caccgtgagg gtgagcaggt cggcggacat ccgcgcgcag 120acggcagcgg tgtcctcccc gtcagtgacc cccgcgtcgc cgtctggcaa gaagaccctc 180cgcataggca cggcggtcat caccggcgcg tcgtccggcc tcggcctcgg cacggcgaag 240gccctcgcgg agacaggcaa g 26144184DNAZea mays 441gtccggcctc ggcctcgcca cggcgaaggc cctcgcggag acaggcaagt ggcacgtcat 60catggcctgc cgcgacttcc tcaa 84442352DNAZea mays 442cggacgcgtg ggctgtcggt gagatcgctt gtggcgacga cggcgcctgt ggccacgccg 60gggtccagca cggcggccaa ggatgggaag aagaccgtgc ggcagggcgt ggtggtgatc 120acgggcgcgt cgtcggggtt gggcctggcg gcggccaagg cgctggcgga gaccggcaag 180tggcacgtgg tgatggcctg ccgcgacttc ctcaaggcgg ccaaggcggc caagggcgcc 240ggcatggcgg acggcagcta caccatcatg cacctggacc tggccttcct cgacagcgtg 300cggcagttcg tggacagctt ccggcgcgcc ggcatgccgc tcgactcgct cg 352443279DNAZea mays 443acgggcgcgt cgtcggggtt gggcctggcg gcggccaagg cgctggcgga gaccggcaag 60tggcacgtgg tgatggcctg ccgcgacttc ctcaaggcgg ccaaggcggc caagggcgcc 120ggcatggcgg acggcagcta caccatcatg cacctggacc tggcctccct cgacagcgtg 180cggcagttcg tggacagctt ccggcgcgcc ggcatgccgc tcgactcgct cgtctgcaac 240gccgccatct accggcccac ggcatagacg ccgacgttc 279444221DNAZea mays 444aaagcgcatc gatctcgctg tcgtcactcc tcgtcaccca gccaaggcgc tggcggagac 60cggcaagtgg cacgtggtga tggcctgccg cgacttcctc aaggcggcca aggcggccaa 120gggcgccggc atggcggacg gcagctacac catcatgcac ctggacctgg cctccctcga 180cagcgtgcgg cagttcgtgg acagcttccg gcgcgccggc a 221445310DNAZea mays 445agtgcagcct cgccgtcgcg gcgaaggaca cggcattcct tagcgtatcc cagaagaagg 60tgcaggcggc gtcgctgtcg gtgagaacgc gggtggcgac gacggcgcct gtggccacgc 120cggggtccag cacggcggcc aaggatggga agaagaccgt gcggcagggc gtggtggtga 180tcacgggcgc gtcgtcgggg ttgggcctgg cggcggccaa ggcgctggcg gagaccggca 240agtggcacgt ggtgatggcc tgccgcgact tcctcaaggc ggccaatgcg gccaagggcg 300ccggcatggc 310446295DNAZea mays 446cccacgcgtc cgcggcgaag gacacggcat tccttagcgt atcccagaag aaggtgcagg 60cggcgtcgct gtcggtgaga acgcgggtgg cgacgacggc gcctgtcgcc acgccggggt 120ccagcacggc ggccaaggat gggaagaaga ccgtgcggca gggcgtggtg gtgatcacgg 180gcgcgtcgtc ggggttgggc ctggcggcgg ccaaggcgct ggcggagacc ggcaagtggc 240acgtggtgat ggcctgccgc gacttcctca aggcggccaa ggcggccaag ggcgc 295447444DNAZea mays 447cggacgcgtg ggcgaacaaa agcgcatcga tctcgctgtc gtcactcctc gtcacccagg 60cacgaacaga ggcaccaccc agcatggccc tgcaggcggc gctcctctca tccaccctct 120catccgtccc caagaagtgc agcctcgccg tcgcggcgaa ggacacggca ttccttagcg 180tatcccagaa ggtcagtgat cagctgcatc tgcatgctgc actcgcagtc acaatgcgct 240tgaattgaac gtgtcactca ctctgtcgtg agcatgccat gcgtgcagaa ggtgcaggcg 300gcgtcgctgt cggtgagagt cacttcgcca tctaccggcc cacggcaagg acgccgacgt 360tcacggcgga cggatacgag atgagcgtcg gcgtcaacca cctgggccac ttcctcctgg 420cgcgcctgct cctggacgac atgc 444448423DNAZea mays 448cccacgcgtc cgcccacgcg tccgcggact cgtgggcttc gccacgaaca aaagcgcatc 60gatctcgctg tcgtcactcc tcgtcaccca gccacgaaca gaggcaccac ccagcatggc 120cctgcaggcg gcgctcctcc catccaccct ctcatccgtc cccaagaagt gcagcctcgc 180cgtcgcggcg aaggacacgg cattccttag cgtatcccag aagaaggtgc aggcggcgtc 240gctgtcggtg agaacgcggg tggcgacgac ggcgcctgtg gccacgccgg ggtccagcac 300ggcggccaag gatgggaaga agaccgtgcg gcagggcgtg gtggtgatca cgggcgcgtc 360gtcggggttg ggcctggcgg cggccaaggc gctggcggag accggcaagt ggcacgtggt 420gat 423449279DNAZea mays 449cgctgtcgtc actcctcgtc acccagccac gaacagaggc accacccagc atggccctgc 60aggcggcgct cctcccatcc accctctcat ccgtccccaa gaagtgcagc ctcgccgtcg 120cggcgaagga cacggcattc cttagcgtat cccacggcgc ggacgccgac gttcacggcg 180gacgggtacg agatgagcgt cggcgtcaac cacctgggcc acttcctcct ggcgcgcctg 240ctcctggacg acatgcagaa gtccgactac acgtcccgc 279450396DNAZea mays 450gacttcgcca cgaacaaaag cgcatcgatc tcgctgtcgt cactcctcgt cacccagcca 60cgaacagagg caccacccag catggccctg caggcggcgc tcctcccatc caccctctca 120tccgtcccca agaagtgcag cctcgccgtc gcggcgaagg acacggcatt ccttagcgta 180tcccagaaga aggtgcaggc ggcgtcgctg tcggtgagaa cgcgggtggc gacgacggcg 240cctgtggcca cgccggggtc cagcacggcg gccaaggatg ggaagaagac cgtgcggcag 300ggcgtggtgg tgatcacggg cgcgtcgtcg gggttgggcc tggcggcggc caaggcgctg 360gcggagaccg gcaagtggca cgtggtgatg gcctgc 396451375DNAZea mays 451cagagtcact tcgccacgaa caaatgcgca tcgatctcgc tgtcgtcact cctcgtcacc 60cagccacgaa cagaggcacc acccagcatg gccctgcagg cggcgctcct cccatccacc 120ctctcatccg tccccaagaa gtgcagcctc gccgtcgcgg cgaaggacac ggcattcctt 180agcgtatccc agaagaaggt gcaggcggcg tcgctgtcgg tgagaacgcg ggtggcgacg 240acggcgcctg tggccacgcc ggggtccagc acggcggcca aggatgggaa gaagaccgtg 300cggcagggcg tggtggtgat cacgggcgcg tcgtcggggt tgggcctggc ggcggccaag 360gcgctggcgg agacc 375452326DNAZea mays 452aacaaaagcg catcgatctc gctgtcgtca ctcctcgtca cccagccacg aacagaggca 60ccacccagca tggccctgca ggcggcgctc ctcccatcca ccctctcatc cgtccccaag 120aagtgcagcc tcgccgtcgc ggcgaaggat caggcattcc ttagcgtatc ccagaagaag 180gtgcaggcgg cgtcgctgtc ggtgagaacg cgggttgcga cgacggcgcc tgttgccacg 240ccggggtcca gcacggcggc caaggatggg aagaagaccg tgcggcaagg cgtggtggtg 300atcacgggcg cgtcgtcggg gttggg 326453338DNAZea mays 453gagtcacttc gccacgaaca aaagcgcatc gatctcgctg tcgtcactcc tcgtcaccca 60gccacgaaca gaggcaccac ccagcatggc cctgcaggcg gcgctcctcc catccaccct 120ctcatccgtc cccaagaagt gcagcctcgc cgtcgcggcg aaggacacgg cattccttag 180cgtatcccag aagaaggtgc aggcggcgtc gctgtcggtg agaacgcggg tggcgacgac 240ggcgcctgtg gccacgccgg ggtccagcac ggcggccaag gatgggaaga agaccgtgcg 300gcagggcgtg gtggtgatca ctggcgcgtc gtcggggt 338454273DNAZea mays 454cttcgccacg aacaaaagcg catcgatctc gctgtcgtca ctcctcgtca cccagccacg 60aacagaggca ccacccagca tggccctgca ggcggcgctc ctcccatcca ccctctcatc 120cgtccccaag aagtgcagcc tcgccgtcgc ggcgaaggac acggcattcc ttagcgtatc 180ccagaagaag gtgcaggcgg cgtcgctgtc ggtgagaacg cgggtggcga cgacggcgcc 240tgtggccacg ccggggtcca gcacggcggc caa 273455296DNAZea mays 455gccacgaaca aaagcgcatc gatctcgctg tcgtcactcc tcgtcaccca gccacgaaca 60gaggcaccac ccagcatggc cctgcaggcg gcgctcctcc catccaccct ctcatccgtc 120cccaagaagt gcagcctcgc cgtcgcggcg aaggacacgg cattccttag cgtatcccag 180aagaaggtgc aggcggcgtc gctgtcggtg agaacgcggg tggcgacgac ggcgcctgtg 240gccacgccgg ggtccagcac ggcggccaag gatgggaaga agaccgtgcg gcaggg 296456314DNAZea mays 456cagagtcagt tcgccacgaa caaaagcgcg tcgatgtcgc tgtcgtcact cgtcgtcacc 60cagccacgaa cagaggcacc acccagcatg gccctgcagg cggcgggtcg tcggatccac 120gctgtcatcc gtccccgaga agtgcagcct cgccgtcgcg gcgaaggtca cggcattcct 180tagcgtatcc cagaagaagg tgcaggcggc gtcggtgtcg gtgagaacgc gggtggcgac 240gacggcgcct gtggccacgc cggggtccag cacagcggcc aaggatggga agaagaccgt 300gcggcagggc gtgg 314457287DNAZea mays 457gagtcacttc gccacgaaca aaagcgcatc gatctcgctg tcgtcactcc tcgtcaccca 60gccacgaaca gaggcaccac ccagcatggc cctgcaggcg gcgctcctcc catccaccct 120ctcatccgtc cccaagaagt gcagcctcgc cgtcgcggcg aaggacacgg cattccttag 180cgtatcccag aagaaggtgc aggcggcgtc gctgtcggtg agaacgcggg tggcgacgac 240ggcgcctgtg gccacgccgg ggtccagcac ggcggccaag gatggga 287458312DNAZea mays 458cagagtcact tcgccacgaa caaaagcgca tcgatctcgc tgtcgtcact cctcgtcacc 60cagccacgaa cagaggcacc acccagcatg gccctgcagg cggcgctcct cccatccacc 120ctctcatccg tccccaagaa gtgcagcctc gccgtcgcgg cgaaggacac ggcattcctt 180agcgtatccc agaagaaggt gcaggcggcg tcgctgtcgg tgagaacgcg ggtggcgacg 240acggcgcctg tggccacgcc ggggtccagc acggcggcca aggatgggaa gaagaccgtg 300cggcagggcg tg 312459321DNAZea mays 459gtcacttcgc cacgaacaaa agcgcatcga tctcgctgtc gtcactcctc gtcacccagc 60cacgaacaga ggcaccaccc agcatggccc tgcaggcggc gctcctccca tccaccctct 120catccgtccc caagaagtgc agcctcgccg tcgcggcgaa ggacacggca ttccttagcg 180tatcccagaa gaaggtgcag gcggcgtcgc tgtcggtgag aacgcgggtg gcgacgacgg 240cgcctgtggc cacgccgggg tccagcacgg cggccaagga tgggaagaag accgtgcggc 300agggcgtggt ggtgatcacg g 321460281DNAZea mays 460cttcgccacg aacaaaagcg cgtcgatctc gctgtcgtca ctcctcgtca cccagccacg 60aacagaggca ccacccagca tggccctgca ggcggcgctc ctcccatcca ccctctcatc 120cgtccccaag aagtgcagcc tcgccgtcgc ggcgaaggac acggcattcc ttagcgtatc 180ccagaagaag gtgcaggcgg cgtcgctgtc ggtgagaacg cgggtggcga cgacggcgcc 240tgtggccacg ccggggtcca gcaggcggcc aaggatggga a 281461314DNAZea mays 461cagagtcact tcgccacgaa caaaagcgca tcgatctcgc tgtcgtcact cctcgtcacc 60cagccacgaa cagaggcacc acccagcatg gccctgcagg cggcgctcct cccatccacc 120ctctcatccg tccccaagaa gtgcagcctc gccgtcgcgg cgaaggacac ggcattcctt 180agcgtatccc agaagaaggt gcaggcggcg tcgctgtcgg tgagaacgcg ggtggcgacg 240acggcgcctg tggccacgcc ggggtccagc acggcggcca aggatgggaa gaagaccgtg 300cggcatggcg tggt 314462351DNAZea mays 462gtccggcaag atgctggcgc aggtggtcag cgaccccagc ctcaccaagt cgggggtgta 60ctggagctgg aacaaggact cggcgtcgtt cgagaaccag ctgtcgcagg aggccagcga 120tccggagaag gccaagaagc tctgggagat cagcgagaag ctcgtggggc ttgcctgagc 180tcgccggcac ggcacagcga catgatggat ctgtcgagca gaggagcttt cgcttcgttg 240tattatgtgt accattagca tccattttgt ttgtttctag aagttggtaa tgaccgtcgg 300agaagagcct gtaattgttc gatcatgtat tgcttacaat ttttttttaa a 351463327DNAZea mays 463gtccggcaag atgctggcgc aggtggtcag cgaccccagc ctcaccaagt cgggggtgta 60ctggagctgg aacaaggact cggcgtcgtt cgagaaccag ctgtcgcagg aggccagcga 120tccggagaag gccaagaagc tctgggagat cagcgagaag ctcgtggggc ttgcctgagc 180tcgccggcac gcgacagcga catgatggat ctgtcgagca gaggagcttt cgcttcgttg 240tattatgtgt accattagca tccattttgt ttgtttctag aagttggtaa tgaccgtcgg 300agaagagcct gtaattgttc gatcatg 327464304DNAZea mays 464ggcctgccgc gacttcctca aggcggccaa ggcggccaag ggcgccggca tggcggacgg 60cagctacacc atcatgcacc tggacctggc ctccttcgac agcgtgcggc agttcgtgga 120cagcttccgg cgcgccggca tgccgctcga ctcgctcgtc tgcaacgccg ccatctaccg 180gcccacggcg cggacgccga cgttcacggc ggacgggtac gagatgagcg tcggcgtcaa 240ccacctgggc cacttcctcc tggcgcgcct gctcctggac gacatgcaga agtccgacta 300cccg 304465285DNAZea mays 465cggcatggcg gacggcagct acaccatcat gcacctggac ctggcctccc tcgacagcgt 60gcggcagttc gtggacagct tccggcgcgc cggcatgccg ctcgactcgc tcgtctgcaa 120cgccgccatc taccggccca cggcgcggac gccgacgttc acggcggacg ggtacgagat 180gagcgtcggc gtcaaccacc tgggccactt cgtcctggcg cgcctgctcc tggacgacat 240gcagaagtcc gactactcgt cccgccgcct cgtcatcctc ggctc 285466147DNAZea mays 466cccacgcgtc cgcacacgcg tccggtggac agcttccggc gcgccggcat gccgctcgac 60tcgctcgtct gcaacgccgc catctaccgg cccacggcgc ggacgccgac gttcacggcg 120gacgggtacg agatgagcgt ccgcgtc 147467280DNAZea mays 467actaaatgcc gaggtgatgg aacttgacct gctctccctc gactcggtcg taaaatttgc 60tgatgcttgg acagctcgta tggcaccgct gcacgtgttg atcaacaatg ctgagctctt 120cgctatagga gaaccccaac atttttccaa ggatggacat gaagaacaca tgcaagtgaa 180ccatcttgca cctgcattac tggcgatgct gcttatacct tcccttctcc gaggttctcc 240cagcagaatt gtaaacgtta attcaatcat gcacagtgta 280468277DNAZea mays 468ctcaaatagc aagctggcac aggtaaaatt cagtagcatg cttcacaaga aaattcctgc 60agaggctggc atcggtgtag tttgcgcttc tcctggaatt gtcgacacga acgttgcaag 120agctcttcct aagattgtcg tagccgcgta ccatttgatt ccctacttca tatttgacgc 180tcaagaaggt tctaggagtg cactgtttgc agcatccgat ccccaagtcc cggaatactg 240cgagacgctc aagtcggagg actggccagt ttgtgcc 277469436DNAZea mays 469ggttctccca gcagaattgt taacgttaat tcaatcatgc acagtgtagg ttttgttgat 60gctgaagatt tgaacttgag aaaacataaa tatagaagtt ggttggcgta ttcaaatagc 120aagttggcac aggtaaaatt tagtagcatg cttcataaga gaattcctgc agaagctggc 180atcagcataa tttgtgcttc

tcctggaatt gtcgacacga atgttacaag agaccttcct 240aagattgttg tagctgcata ccattttctt ccctacttca tattcgatgg tcaagaaggt 300tctaggagtg cactgtttgc agcatgtgac ccccaagttc cagagtactg tgagatgctc 360aagtcggaag actggccagt ctgtgcttgc attaactacg actgtaatcc gatgaacgcg 420tctgaagaag cgcaca 436470335DNAZea mays 470gtagaattta gtagcatgct tcataagata attcctgcag aagctggcat cagcataatt 60tgtgcttctc ctggaattgt cgacacgaat gttacaagag accttcctaa gattgttgta 120gctgcatacc gttttcttcc ctacttcata ttcgatggtc aagaaggttc taggagtgca 180ctgtttgcag catgtgaccc ccaagttcca gagtactgtt gagatgctca agtcggaaga 240ctggccagtc tgtgcttgca ttaactacga ctgtaatccg atgaacgcgt ctgaagaagc 300gcacagcttg ataccttcgc agctggtctg ggaga 335471343DNAZea mays 471gtaaaatgta gtagcatgct tcataagaga attcctgcag aagctggcat cagcataatt 60tgtgcttctc ctggaattgt cgacacgaat gttacaagag accttcctaa gattgttgta 120gctgcatacc gttttcttcc ctacttcata ttcgatggtc aagaaggttc taggagtgca 180ctgtttgcag catgtgaccc ccaagttcca gagtactgtg agatgctcaa gtcggtagac 240tggccagtct gtgcttgcat taactacgac tgtaatccga tgaacgcgtc tgaagaagcg 300cacagccttg aaacctcgca gctggtctgg gagaagcgct cga 343472262DNAZea mays 472gtaaaattta gtagcatgct tcataagata attcctgcag aagctggcat cagcataatt 60tgtgcttctc ctggaattgt cgacacgaat gttacaagag accttcctaa gattgttgta 120gctgcatacc gttttcttcc ctacttcata ttcgatggtc aagaaggttc taggagtgca 180ctgtttgcag catgtgaccc ccaagttcca gagtactgtg agatgctcaa gtcggaagac 240tggccagtct gtgcttgcat ta 262473256DNAZea mays 473gcttcataag agaattcctg cagaagctgg catcagcata atttgtgctt ctcctggaat 60tgtcgacacg aatgttacaa gagaccttcc taagattgtt gtagctgcat accgttttct 120tccctacttc atattcgatg gtcaagaagg ttctaggagt gcactgtttg cggcatgtga 180cccccaagtt ccagagtact gtgagatgct caagtcggaa gactggccag tctgtgcttg 240cattaactac gactgt 256474208DNAZea mays 474gcttcataag agaattcctg cagaagctgg catcagcata atttgtgctt ctcctggaat 60tgtcgacacg aatgttacaa gagaccttcc taagattgtt gtagctgcat accgttttct 120tccctacttc atattcgatg gtcaagaagg ttctaggagt gcactgtttg cggcatgtga 180cccccaagtt ccagagtact gtgagatg 208475338DNAZea mays 475gtatgattta gtagcatgct gcataagaga gttcctgcag aagctggcat cagcataatt 60tgtgcttctc ctggaattct cgacacgaat gttacgagaa tccttcctaa gattgttgta 120gctgcatacc gttgtcttcc ctacttcata ttcgatggtc aacaaggttc taggagtgca 180ctgtctgcag catgtgaccc ccaagttcca gagtactgtg agatgctcaa gtcggaagac 240tggccagtct gtgcttgcat taactacgac tgtaatccga tgaacgcgtc tgaagaagcg 300cacagccttg aaacctcgca gctggtctgg gagaagac 338476248DNAZea mays 476gattgatgct gaagatttca acttgagaaa acataaatat agaagttggt tggcgtattc 60aaatagcaag ttggcacagg taaaatttag tagcatgctt cataagagaa ttcctgcaga 120agctggcatc agcataattt gtgcttctcc tggaattgtc gacacgaatg ttacaagaga 180ccttcctaag attgttgtag ctgcatacgg tttcccccaa atcaaaatcg atggtcaaga 240aggttcta 248477341DNAZea mays 477gagatcttcc taagattgtc gtagccgcgt accatttgat tccctacttc atatttgacg 60ctcaagaagg ttctaggagt gcactgtttg cagcatccga tccccaagtc ccggagtact 120gcgagacgct caagtcggag gactggccag tttgtgcctg cattaactat gactgtagtc 180cgatgaatgc gtctgaagaa gcgcacaatc tggagacctc gcagctggtc tgggagaaga 240cactggagat ggtcggcctt ccgccggatg ccctggagaa gctcatcgcc ggagaatcag 300ttcagtgccg ttacggacaa caggatacaa cttaactttt t 341478383DNAZea mays 478gtgcactgtt tgcagcatcc gatccccaag tcccggaata ctgcgagacg ctcaagtcgg 60aggactggcc agggggtgcc tgcattaact atgactgtag tccgatgaat gcgtctgaag 120aagcgcacaa tcttgagacc tcgcagctgg tctgggagaa gacactggag atggtcggcc 180ttccgccgga tgccctggag aagctcatcg ccggagaatc agttcagtgc cgttacggac 240aacaggatac aactttttag ttagcagttt agaggtggtt tgttcggttg ttatgtcatt 300ttgatcctaa atttgcaggg aggaaaacac agggaaagga gaaaaagaat ttgttgacag 360ctacccaatc ttggctcttt tct 383479166DNAZea mays 479ggaggactgg ccattttgtg cctgcatgaa ctatgactgt agtccgatga atgcgtctta 60caggagcgca caatcttgag acctcgcagc tggtctggga gaagacactg gagatggtcg 120gcgttccgcc ggatgccctg gagaagctca tcgccggaga atcagt 166480382DNAZea maysunsure at all n locations 480agtgaggagt ngcttccaaa actgatgcat gnantcatgc aatacgcatt ccggtcgacc 60actcgtaccc tggtaaaccc gaaggattgg atctgattat ccgctattct tgtgtccctt 120acgcttggag cacgatggca gtatgatcat aaaccggatg aaggaaccgc cgaacggaaa 180cttctataag cctgcataaa cccgatagat tggatctgat tatcccttat tcttgagatc 240tttagttaga gttttccctt ctgtagggct aaaaccacgt gcagcttcat gatatatcct 300gcctctgtac aatcgtgaac aaatattacg tattaatgct ctatctgcct gtattatata 360tgctgctttt tgcccatgtg aa 382481358DNAZea mays 481cctgcataaa cccgaaggat tggatctgat tagccgttat tcttgtgtcc cttccgcttg 60cagcacgatg gcagtatgat cataaaccgg aagaaggaac cgaggaatgg aaacttctgg 120aagcctgcat aaacccgaag gattggatct gattagccgt tattcttgag atcttttgtt 180agagttttcc cttctgtagg gctaagacca cgtgcagttt cattatatat tttgcatctg 240tagaatcgtg aataaatatg atgtagtaat gctgtagctg tctgtatcta tctgctgttt 300tttccccatg tgaatgagag aaccattggc ttctgtatta cgaaggattc aggtttct 358482275DNAZea mays 482accggaagaa ggaaccgagg aatggaaact tctggaagcc tgcataaacc cgaaggattg 60gatctgatta gccgtcattc ttgagatctt ttgttagagt tttcccttct gtagggctaa 120gaccacgtgc agtttcatta tttctttttg catctgtaga atcgtgaata aatatgatgt 180agtaatgctg tagctgtttg tatctatctg ctgttttttc cccatgtgaa tgagtgaacc 240attggcttct gtatttacga aggattcagg tttct 275483335DNAZea mays 483cttgaagagg acgtgaagca tttccattct gttcaaaagc aagcatgtga taaatttgat 60ccaagttttc acccaagatt caaaaaatgg tgtgatgatt atttctatat taagcaccgt 120aatgagcggc gtgggctagg tggaatattt tttgatgacc ttaatgatta cgatcaagaa 180atgcttctca actttgctac agaatgtgcg gactctgtac ttcctgcgta cataccgatc 240atagaacggc ggaagaacac tccgttcaat gaggagcaca gggcatggca gcaattgcgg 300agaggtcgtt atgtggagtt caaccttgtc tacga 335484475DNAZea mays 484caagaaatgc ttctcaactt tgctacagaa tgtgcggact ctgtacttcc tgcgtacata 60ccgatcatag aacggaggaa gaacactccg ttcaacgagg agcacagggc atggcagcaa 120ttgcggagag gtcgttatgt ggagttcaac cttgtctacg accgtggtac aacatttggc 180ctaaagactg gaggaaggat tgagagcata cttgtgtccc ttccacttac agcacgatgg 240cagtatgatc ataaaccgga agaaggaacc gaggaatgga aacttctgga agcctgcata 300aacccgaagg attggatctg attagccgtt attcttgaga tcttttgtta gaagtttccc 360ttctgtaggg ctaagaccac gtgcagtttc attatatatt ttgcatctgt agaatcgtga 420ataaatatga tgtagtgatg ttgtagctgt ttggatctat ctgctggttt ttccc 475485329DNAZea maysunsure at all n locations 485atcaagaaat gcttctcaac tttgctacag aatgtgcgga ctctgtactt cctgcgtaca 60taccgatcat agaacggagg aagaacactc cgttcaacga ggagcacagg gcatggcagc 120aattgcggag aggtcgttat gtggagttca accttgtcta cgaccgtggt acaacatttg 180gcctaaagac tggaggaagg attgagagca tacttgtgtc ncttccactt acagcacgat 240ggcagtatga tcatanaccg gaagaaggaa ccgacgaatg ganacttctg gaagcctgca 300tagacccgaa ggattggatc tgattagcg 329486270DNAZea mays 486caagattcaa aatatggtgt gatgattatt tctatattaa gcaccgtaat gagcggcgtg 60ggctaggtgg aatatttttt gatgacctta atgattacga tcaagaaatg cttctcaact 120ttgctacaga atgtgcggac tctgtacttc ctgcgtacat accgatcata gaacggagga 180agaacactcc gttcaacgag gagcacaggg catggcagca attgcggaga ggtcgttatg 240tggagttcaa ccttgtctac gaccgtggta 270487256DNAZea mays 487cgcggcgtgg gctaggtgga atattttttg atgaccttaa tgattacgat caagaaatgc 60ttctcaactt tgctacagaa tgtgcggact ctgtacttcc tgcgtacata ccgatcatag 120aacggaggaa gaacactccg ttcaacgagg agcacagggc atggcagcaa ttgcggagag 180gtcgttatgt ggagttcaac cttgtctacg accgtggtac aacatttggc ctaaagactg 240gaggacggat tgacag 256488247DNAZea mays 488cttaatgatt acgatcaaga aatgcttctc aactttgcta cagaatgtgc ggactctgta 60cttcctgcgt acataccgat catagaacgg cggaagaaca ctccgttcaa tgaggagcac 120agggcatggc agcaattgcg gagaggtcgt tatgtggagt tcaaccttgt ctacgaccgt 180ggtaccacat ttggcctaaa gactggagga aggattgaga gcatacttgt gtcccttccg 240cttacag 247489236DNAZea mays 489cccacgcgtc cgctccgttc aatgaggagc acagggcatg gcagcaattg cggagaggtc 60gttatgtgga gttcaacctt gtctacgacc gtggtaccac atttggccta aagactggag 120gaaggattga gagcatactt gtgtcccttc cgcttacagc acgatggcag tatgatcata 180aaccggaaga aggaaccgag gaatggaaac ttctggaagc ctgcataaac ccgaag 236490430DNAZea mays 490gggggaggcc gccaagaacg gggccgccgc cgcggatggc cacaagcctg ggccggtggc 60attcttcgcc gcggggatta gttcggtgct tcaccccaag aacccatttg ctccaacatt 120gcattttaac taccgttact ttgagacgga tgcaccaaaa gatgcacctg gtgcaccaag 180acaatggtgg ttcggcggtg gtactgactt gactccttca tatatcattg aagaggatgt 240gaagcatttc cattctgttc aaaagcaagc atgtgataaa tttgatccaa gttttcaccc 300aagattcaaa aaatggtgtg atgattattt ctatattaag caccgtaatg agcggcgtgg 360gctaggtgga atattttttg atgaccttaa tgattacgat caagaaatgc ttctcaactt 420tgctacagaa 430491304DNAZea mays 491gggccgccgc cgcggatggc cacaagcctg gccccgtgcc attcttcgcc gcggggatta 60gttcggtgct tcaccccaag aacccatttg ctccaacatt gcattttaac taccgttact 120ttgagacgga tgcaccaaaa gatgcacctg gtgcaccaag acaatggtgg ttcggcggtg 180gtactgactt gactccttca tacatcattg aagaggacgt gaagcatttc cattctgttc 240aaaagcaagc atgtgataaa tttgatccaa gttttcaccc aagattcaaa aaatggtgtg 300atga 304492307DNAZea mays 492ggaggccgcc aagaacgggg ccgccgccgc ggatggccac aagcctggcc ccgtgccatt 60cttcgccgcg gggattagtt cggtgcttca ccccaagaac ccatttgctc caacattgca 120ttttaactac cgttactttg agacggatgc accaaaagat gcacctggtg caccaagaca 180atggtggttc ggcggtggta ctgacttgac tccttcatac atcattgaag aggacgtgaa 240gcatttccat tctgttcaaa agcaagcatg tgataaattt gatccaagtt ttcacccaag 300attcaaa 307493173DNAZea mays 493gcacgagaaa agatgcacct ggtgcaccaa gacaatggtg gttcggcggt ggtactgact 60tgactccttc atacatcatt gaagaggacg tgaagcattt ccattctgtt caaaagcaag 120catgtgataa atttgatcca agttttcacc caagattcaa aaaatggtgt gat 173494118DNAZea mays 494gttactttga gacggatgca ccaaaagatg cacctggtgc accaagacaa tggtggttcg 60gcggaggtac tgacttgact ccttcataca tcattgaaga ggacgtgaag catatcca 118495304DNAZea mays 495agaagccgca aaaactgccc tggaccgagg tggctacgat gggctgttcc taggagggaa 60ctatgttgca ggagttgacc tgggcagatg cgttgagggc gcgtatgaaa gtgcctcgca 120aatatctgac ttcttgacca agtatgccta caagtgatga aagaagtgga gcgctacttg 180ttaattgttt atgttgcata gatgaggtgc ctacgggaaa aaaaagcttt aatagtattt 240tttattctta ttttgtaaat tgcatttctg ttcttttttc tgtcattaat tacttatatt 300ttag 304496295DNAZea mays 496gagggaacta tgttgcagga gttgccctgg gcagatgcgt tgagggcgcg tatgaaagtg 60cctcgcaaat atctgacttc ttgaccaagt atgcctacaa gtgatgaaag aagtggagcg 120ctacttgtta atcgtttatg ttgcatagat gaggtgcctc cggggaaaaa aagcttgaat 180agtatttttt attcttattt tgtaaattgc atttctgttc ttttttctat cagtaattag 240ttatatttta gttctgtagg agattgttct gttcactgcc cttcaaaaga atttt 295497305DNAZea mays 497cgttcttcga tctcatgagc atcccaggga agctcagggc cggtctaggc gcgcttggca 60tccgcccgcc tcctccaggc cgcgaagagt cagtggagga gttcgtgcgc cgaacttcgt 120gctgaggtct tcgagcgcct cattgagcct ttctgctcag gtgtctatgc tggtgatcct 180tctaagctca gcatgaaggc tgcatttggg aaggtttggc ggttggaaga aactggaggt 240agtattattg gtggaaccat caagacaatt caggagagga gcaagaatcc aaaaccactg 300aggga 305498270DNAZea mays 498ggacctggcc gcccgcctcc tccaggccgc gaagagtcag tggaggagtt cgtgcgccgc 60aatcttggtg ctgaggtctt cgagcgcctc attgagcctt tctgctcagg tgtctatgct 120ggtgatcctt ctaagctcag catgaaggct gcatttggga aggtttggcg gttggaagaa 180actggaggta gtattattgg tggaacatca agacaattca ggagaggagc aagaatccaa 240aaccactgag ggatgcccgc cttccgaagc 270499423DNAZea mays 499atccaaagga agcaattaga aaagaatgct taattgatgg ggagctccag ggcgttgggc 60agttgcatcc acgtagtcaa ggagttgaga cattaggaac aatatacagt tcctcactct 120ttccaaatcg tgctcctgac ggtagggtgt tacttctaaa ctacatagga ggtgctacaa 180acacaggaat tgtttccaag actgaaagtg agctggtcga agcagttgac cgtgacctcc 240gaaaaatgct tataaattct acagcagtgg accctttagt ccttggtgtt cgagtttggc 300cacaagccat acctcagttc ctggtaggac atcttgatct tctggaagcc gcaaaagctg 360ccctggaccg aggtggctac gatgggctgt tcctaggagg gaactatgtt gcaggagttg 420ccc 423500314DNAZea mays 500cacgcccctg ccggccatcg gggtgccgtt cgatatctcg gactccaagg ggcccgtgat 60ccaatcgcca gtacggtcca aagagcaggt gagggagctc gtccccatcg accttgatat 120gctccagttc gtcggggagt cactaaagat tctgcgaaat gagattgatg gaaaagctgc 180tttgctagga tttgtggggg ccccatggac aattgcaact tacattgttg aaggggggat 240gaccaatacg tacacaaata taaagagcat gtgccataca gctccagatg tcttgaaggg 300tcttctctct cact 314501287DNAZea mays 501gaaggaggtt catcaaagaa ctttacattg attaagaaaa tggccttctc agaaccagcg 60attctacaca atttgctaca gaagttcaca acatcaatgg ctaactatat taaataccaa 120gcggacaatg gggcgcaggc tgtccaaatt ttcgattcat gggctactga actcagcccg 180actgattttg aggagtttag cctgccttat ctaaagcaga tagtggatag tgttagggaa 240acacatccta acttgcctct gatactctac gcaagtggat ctggggg 287502272DNAZea mays 502gtccagtgta tacagatatt tgattcatgg ggtggacagc ttccacctca tgtatgggag 60cagtggtcaa aaccatatat caaacaggag ttgatgttat tgggcttgac tggacagtgg 120acactactga tggaaggtgg cgccttggta atggcattag tgtacaaggg aatgtggatc 180cagcattttt gttctcacca ttaccagtac tgactgatga aattcataga gttgtgaaag 240cagctggtcc aaaaggtcat accttaatct gg 272503407DNAZea mays 503agggcagagg gcaggaaaag attgggatct aacacagcag tccaagggaa cgtggatcct 60ggtgttcttt ttggatccaa agagtttata agcaggcgga tttacgacac tgtgcagaag 120gctggcaatg ttggacatgt actgaacctt ggccatggca tcaaggttgg aactccggag 180gaaaatgttg ctcacttctt cgaggtcgca aaagggatca gatactaaag aaccttgcat 240ggttctttcc tttctccaaa tcggcagaag ttgtagagtc ggcggtcgag gatagatgca 300gaaagccatg tgcagtatag agtccctgaa aacatttttg tgactgattc tgtctgtcgc 360aattcaagtt ccggtttcaa tgtgatattg taagcagatt tgagacg 407504418DNAZea mays 504agcaagtgaa ggccaggttg cgggaggcag gcctggcacc agtgcccatg atcatctttg 60ctaaggatgg gcattttgcc ctggaggagc tggcccaagc tggctatgag gtggttgggc 120ttgactggac agtggcccca aagaaagccc gggagtgtgt ggggaagacg gtgacattgc 180agggcaacct ggacccctgt gccttgtatg catctgagga ggagatcggg cagttggtga 240agcagatgct ggatgacttt ggaccacatc gctacattgc caacctgggc catgggcttt 300atcctgacat ggacccagaa catgtgggcg cctttgtgga tgctgtgcat aaacactcac 360gtctgcttcg acagaactga gtgtatacct ttaccctcaa gtaccactaa cacagatg 418505508DNAZea maysunsure at all n locations 505cgagctggct gccattagag ccttcgcaac agaaataant agctaccgtc agccaccggt 60tccggtaatt cgccggggga ggacccaccg cgtgccgcga gcggctgcaa ccacctactc 120attgcgtttt caatggcaac aacgtgtacg tcggtctcgg tgccgtgcac cttcctcttg 180cgcggcaggt ccgcccgcac catgcccaga cgcaagcagc tcacggccgt ccgctgcagc 240gccgtcagac aggccgtagt ggaagaggcc tcgcccggga ccgcggacga tccgctgctg 300gtgagcgcaa tcagagggac gaaggtcgag aagccacccg tatggctcat gaggcacgcc 360gggaggtaca tgaagagcta ccaattgctc tgcgagcggc atccttcgtt ccgtgaaaga 420tcagaaaatg tcgacctagt tgttgagatc tctttgcaac catggaaggt tttcaagcct 480gaaggaatca tcttggtctc ggacattc 508506387DNAZea mays 506cccacgcgtc cgcccactcg tccgaaattt tcgattcatg ggctactgag ctcagcccgg 60ctgattttga ggagtttagc ctgccttatc taaagcagat agtggatagt gttagggaaa 120cacatcctaa cttgcctctg atactctacg caagtggatc tgggggcttg ctggagaggc 180ttcctttgac aggtgttgat gttgtcagct tggactggac ggtcgatatg gcagagggca 240ggaaaagatt gggatctaac acagcagtcc aagggaacgt ggatcctggt gttctttttg 300gatccaaaga gtttataagc aggcggattt acgacactgt gcagaaggct ggcaatgttg 360gacatgtact gaaccttggc catggca 387507288DNAZea mays 507gccgctgctg gtgagcgcaa tcagaaggag gaaggtcgag aagccacccg tctggctcat 60gaggcaggcc gggaggtaca tgaagagcta ccaattgctc tgcgagcggt atccttgttc 120cgtgaaagat cagaaaatgt cgacctagtt gttgagatct ctttgcaacc atggaaggtt 180ttcaagcctg atggagtcat cttgttctcg gacatcctta ctccacttcc tgggatgaac 240ataccttttg acattgtgaa gggaaaaggt ccagtgatct atgatcca 288508409DNAZea mays 508gtccgcgagc gctgcagcac ctcggatccc gccccaatgg caacagcgtg tccgccgctc 60tcgctgccgt ccacctccct cttccgcggc aggtccgccc gcgccgggcc cagacgcagg 120cagctcacgg ccgtccgctg cagcgccgtc ggagaggcgg tagtggagga ggcctcgccc 180gggacggcgg aagagccgct gctggtgagc gcaatcagag ggaggaaggt cgagaggcca 240cccgtctggc tcatgaggca ggccgggagg tacatgaaga gctaccaatt gctctgcgag 300cggtatcctt cgttccgtga aagatcagaa aatgtcgacc tagttgttga gatctctttg 360caaccatgga aggttttcaa gcctgatgga gtcatcttgt tctcggaca 409509407DNAZea mays 509agccaagtcg tcgcctcccc gacccaacgt tttgaccccc ttgcccgtcc gcgagcgctg 60cagcacctgg gatcccgccc caatggcaac agcgtgtccg ccgctctcgc tgccgtccac 120ctccctcttc cgcggcaggt ccgcccgcgc cgggcccaga cgcaggcagc tcacggccgt 180ccgctgcagc gccgtcggag aggcggtagt ggaggaggcc tcgcccggga cggcggaaga 240gccgctgctg gtgagcgcaa tcagagggag gaaggtcgag aggccacccg tctggctcat 300gaggcaagcc gggaggtaca tgaagagcta ccaattgctc tgcgagcggt atccttcgtt 360ccgtgaaaga tcagaaaatg tcgacctagt tgttgagatc tctttgc 407510275DNAZea mays 510taaagattct gcgaaatgag attgatggaa aagctgcttt gctaggattt gtgggggccc 60catggacaat tgcaacttac attgttaaag gggggatgac

caacacatac acaaatataa 120agaacatgtg ccatacagct cccgatgtct taggtgtctt ctatctcatc ttgcagtagc 180gatatctgac tatatcattt accaagttaa ctccggggcc cagtgtatac agatatttga 240ttcatggggc ggacaacttc cacctcatgt gtggg 275511266DNAZea maysunsure at all n locations 511tgccaagagc cgggccaagg ctgcgctcca cggccgtccg ggtcagcagc gagcaggagg 60cggcggcggc cgtcnaggcg ccgtccggga ggaccatcga ggagtgcgag gccgacgccg 120tcgctgggaa gttccctgct cccccgccgc tggttaggcc gaagcgcctg aaggaacgcc 180ggagatcagg ccccttgaca tggcaaagcg cccccgtcgc aaccgcaaat cacctgctct 240tagggctgca ttccaggaga cgagca 266512293DNAZea mays 512gccgtacttg gacattatcc gactgcttcg ggatcattca gccctaccga ttgctgctta 60ccaggtctcg ggcgagtact cgatgatcaa agccggcggg gccctgggca tggtggacga 120gcagaaggtg atgatggagt cgctcatgtg cctgcgcgag ccggcgccga cgtcatcctg 180acctacttcg cccgtcacgc cgccgcggtg ctgtgcggca tggggcccaa gtaggaggcg 240aggcccgccc gccattcctg ccctgcactg tcattgtgga gttgagcgat gag 293513279DNAZea mays 513actagattca catccaagat ttggagataa gaagacgtac cagatgaacc cagctaacta 60cagagaagcc ctcatagaaa ccgcatcgga cgaggcagaa ggagccgaca ttctgctagt 120gaaaccggga ttgccgtact tggacattat ccgactgctt cgggatcatt cagccctacc 180gagtgctgct taccaggtct cgggcgagta ctcgatgatc agagccggag gggccctggg 240catggtggac gagcataagg tgatgatgga gtcgctcat 279514287DNAZea mays 514cggacgcgtg gggttcattt tatggccctt ccgagaagct ttagattcaa atccaagatt 60tggagataag acgacgtacc agatgaaccc agccaactac agagaagccc tcatagaaac 120cgcagcggac gaggcagaag gagccgacat tctgctagtg aaaccgggat tgccgtactt 180ggacatcatc cgactgcttc gggatcattc agccctaccg attgctgctt accaggtctc 240gggcgagtac tcgatgatca aagccggcgg ggccctgggc atggtgg 287515427DNAZea mays 515ctttgtgctc ccattgttta tccatgaagg agaagaagat gctcctatcg gagctatggc 60agggtgctat aggcttgggt ggaggcacgg gctgcttgac gaggtttaca aggcccgcga 120tgttggtgtt aatagtttcg ttctctttcc taaagttccc gatgcattga agtctccaac 180aggagatgaa gcgtacaacg ataatggtct ggttccacgt acaatccgct tgctcaagga 240caagttccct gatattgtta tctacacaga cgtcgcgtta gacccttatt catctgatgg 300tcatgatggt attgtgaggg aagatggtgt aattatgaat gatgaaacag tttatcagtt 360gtgcaaacag gctgtttcac aggctcgtgc cggtgctgat gttgtcagcc ctagtgacat 420gatggat 427516303DNAZea mays 516cccacgcgtc cgcaaggccc gcgatgttgg tgttaatagt ttcgttctct ttcctaaagt 60tcccgatgca ttgaagtctc caacaggaga tgaagcgtac aacgataatg gtctggttcc 120acgtacaatc cgcttgctca aggacaagtt ccctgatatt gttatctaca cagacgtcgc 180gttagaccct tattcatctg atggtcatga tggtattgtc agggaagatg gtgtaattat 240gaatgatgaa acagtttatc agttgtgcaa acaggctgtt tcacaggctc gtgccggtgc 300tga 303517277DNAZea mays 517cttattcatc tgatggtcat gatggtattg tgagggaaga tggtgtaatt atgaatgatg 60aaacagttta tcagttgtgc aaacaggctg tttcacaggc tcgtgccggt gctgatgttg 120tcagccctag tgacatgatg gatggccgga ttggagcact tcgctctgct ctggacgccg 180agggcttcca tgatgtctcc attatgtcct acaccgcaaa gtatgccagt tcattttatg 240gccctttccg agaagcttta gattcaaatc caagatt 277518300DNAZea mays 518cccacgcgtc cgcaaggccc gcgatgtagg tgttaatagt ttcgttctct ttcctaaagt 60tcccgatgca ttgaagtctc caacaggaga tgaagcgtac aacgataatg gtctggttcc 120acgtacaatc cgcttgctca aggacaagtt ccctgatatt gttatctaca cagacgtcgc 180gttagaccct tattcatctg atggtcatga tggtattgtt agggaagatg gtgtaattat 240gaatgatgaa acagtttatc agttgtgcaa acaggctgtt tcacaggctc gtgccggtgc 300519306DNAZea mays 519cccacgcgtc cgcccacgcg tccgcccacg cgtccgccca cgcgtccggg acaagttccc 60tgatattgtt atctacacag acgtcgcgtt agacccttat tcatctgatg gtcatgatgg 120tattgtgagg gaagatggtg taattatgaa tgatgaaaca gtttatcagt tgtgcaaaca 180ggctgtttca caggctcgtg ccggtgctga tgttgtcagc cctagtgaca tgatggatgg 240ccggattgga gcacttcgct ctgctctgga cgccgagggc ttccatgatg tctccattat 300gtccta 306520391DNAZea mays 520acgaacgcgt gggcggacgc gtgggcggac gcgtgggaga acgcgtgggc ggacgcgtgg 60gtgaaggaga agaagatgct cctatcggag ctatgccagg gtgctatagg cttgggtgga 120ggcacgggct gcttgacgag gtttacaggg gcgcgcgatg ttggtgttaa tagttttgtt 180ctctttccta aagttcccga tgcattgaag tctccaacag gagatgaagc gtacaacgat 240aatggtctgg ttccacgtac aatccgcttg ctcaaggaca agttccctga tattgttatc 300tacacagacg tctctttttt ttcttagtca tctgatggtc actatggtat tgttacggaa 360gatggggtaa ttatgaatga tgaaacactt t 391521191DNAZea mays 521agatgctcct atcggagcta tgccagggtg ctataggctt gggtggaggc acgggctgct 60tgacgaggtt tacaaggccc gcgatgttgg tgttaatagt ttcgttctct ttcctaaagt 120tcccgatgca ttgaagtctc caacaggaga tgaagcgtac aacgataatg gtctggttcc 180acgtacaatt c 191522128DNAZea mays 522gttagaccct tattcatctg atggtcatga tggtattgtg agggaagatg gtgtaattat 60gaatgatgaa acagtttatc agttgtgcaa acaggctgtt tcacaggctc gtgccggtgc 120tgatgttg 128523301DNAZea mays 523gcagcttctc cgtgctgctg cgtctcctcc tcatcgtcct ctccagtgtc cagctcggcc 60atggcgttca ccgtctcctt ctcccccgcc aacgttcaga tgctccaggc taggagtggc 120cacggccacg ccacctttgg aagctgttcc gccgtgccaa gagccgggcc aaggctgcgc 180tccacggccg tccgggtcag cagcgagcag gaggcggcgg cggccgtcag ggcgccgtcc 240gggaggacca tcgaggagtg cgaggccgac gccgtcgctg ggaagttccc tgctcccccg 300c 301524323DNAZea mays 524caggattagc agcttctccg tgctgctgcg tctcctcctc atcgtcctct ccagtgtcca 60gctcggccat ggcgttcacc gtctccttct cccccgccaa cgttcagatg ctccaggcta 120ggagtggcca cggccacgcc acctttggaa gctgttccgc cgtgccaaga gccgggccaa 180ggctgcgctc cacggccgtc cgggtcagca gcgagcagga ggcggcggcg gccgtcaggg 240cgccgtccgg gaggaccatc gaggagtgcg aggccgacgc cgtcgctggg aagttccctg 300ctcccccgcc gctggttagg ccg 323525252DNAZea mays 525cagattagca gcttctccgt gctgctgcgt ctcctcctca tcgtcctctc cagtgtccag 60ctcggccatg gcgttcaccg tctccttctc ccccgccaac gttcagatgc tccaggctag 120gagtggccac ggccacgcca cctttggaag ctgttccgcc gtgccaagag ccgggccaag 180gctgcgctcc acggccgtcc gggtcagcag cgagcaggag gcggcggcgg ccgatcaggc 240gccgtccggg ag 252526304DNAZea maysunsure at all n locations 526cacaggatta gcagcttctc cgtgctgctg cgtctcctcc tcatcgtcct ctccagtgtc 60cagctcggcc atggcgttca ccgtctcctt ctcccccgcc aacgttcaga tgctccaggc 120taggagntgg cacggccacg ccacctttgg aagctgttcc gccgtgccaa gagccgggcc 180aaggctgcgc tccacggccg tccgggtcag cagcgagcag gaggcggcgg cggccgtcag 240ggcgccgtcc gggaggacca tcgaggagtg cgaggccgac gccgtcgctg ggaagttccc 300tgct 304527295DNAZea maysunsure at all n locations 527cacaggatta gcagcttctc cgtgctgctg cgtctcctcc tcatcgtcct ctccagtgtc 60aagctcggcc atggcgttca ccgtctcctt ctcccccgcc aacgttcaga tgctccaggc 120taggagtggc cacggccacg ccacctttgg aagctgttcc gccgtgccaa gagccgggcc 180aaggctgcgc tccacggccg tccgggtcag cagcgagcag gaggcggcgg cggccgtcag 240gcgccgtccg ggaggaccat cgaggantcg aagccgacgc cgtgctggga nnttc 295528239DNAZea mays 528ccacgcgtcc gcagattagc agcttctccg tgctgctgcg tctcctcctc atcgtcctct 60ccagtgtcca gctcggccat ggcgttcacc gtctccttct cccccgccaa cgttcagatg 120ctccaggcta ggagtggcca cggccacgcc acctttggaa gctgttccgc cgtgccaaga 180gccgggccaa ggctgcgctc cacggccgtc cgggtcagca gcgagcagga ggcggcggc 239529302DNAZea mays 529acaggattag cagcttctcc gtgctgctgc gtctcctcct catcgtcctc tccagtgtcc 60agctcggcca tggcgttcac cgtctccttc tcccccgcca acgttcagat gctccaggct 120aggagtggcc acggccacgc cacctttgga agctgttccg ccgtgccaag agccgggcca 180aggctgcgct ccacggccgt ccgggtcagc agcgagcagg aggcggcggc ggccgtcaag 240gcgccgtccg ggaggaccat cgaggagtgc gaggccgacg ccgtcgctgg gaagttccct 300gc 302530242DNAZea mays 530gccacgggtc cgcagtatta gcagcttctc cgtgctgctg cgtctcctcc tcatcgtcct 60ctccagtgtc cagctcggcc atggcgttca ccgtctcctt ctccccagcc aacgttcaga 120tgctccaggc taggagtggc cacggccacg ccacctttgg aagctgttcc gccgtgccaa 180gagccgggcc aaggctgcgc tcaacggccg tccgggtcag cagcgagcag gaggcggcgg 240cg 242531255DNAZea mays 531cccacgcgtc cgaccacgcg tccgcggacg ctggccccgg cgatgatgga cctctccagt 60gtccagctcg gccatggcgt tcaccgtctc cttctccccc gccaacgttc agatgctcca 120ggctaggagt ggccacggcc acgccacctt tggaagctgt tccgccgtgc caagagccgg 180gccaaggctg cgctccacgg ccgtccgggt cagcagcaag caaaaggcgg cgacggacgt 240caggcggcgt cccgg 255532280DNAZea mays 532ctcttttgac gacatggttg agatgggcaa agatgctggc catgagctga aggcaaaggc 60tgggcctggc ttctttgata gcttgcaatg aaaagaatga gcgaccatga gcaatttcaa 120ttgtcactct tttggttaga aacagagggc ccaagtagag tgtggagagg tttgtttttg 180tttcttcttt ctcctgctaa ttctgctaga gaagggtgta cctggtgtag tggtgagccg 240agtcatcagg tcgcgggttc gaagcatcca gtctccgtat 280533325DNAZea mays 533aaacacgcgt ccgcggacgc tggggacacg gttaaggaaa ctcaaggaag gagatgtgtc 60tgctacattg taggcgcagg ctgagattaa ggcggctaaa tatggcagaa aatgcaacag 120ctgtactatc agtggaagaa atgcttccgg cagttgccca aggtgctatt ggaatcgctt 180gccgaagcaa cgatgacaaa atgatggagt atctgtcctc gttgaaccac gaggatacca 240gactagctgt cacatgcgaa agagaattct tggcagttct tgatggcaac tgccgaactc 300caattgcggc ctatgcttac cgtga 325534282DNAZea mays 534tgcattcata tgcttgactg caaattctct cgcggagctt cctgctggca gtgttggtgg 60aagtgcttcc ttgcctagac aatctcacat tctctacaga tatccatcac tgaaagtagt 120taacttcaga ggaaatgttc agacacggtt aaggaaactc actgaaggag atgtgtctgc 180tacattgttg gcgctggctg gattaaggca gctaaatatt gcagaaaatg caacagctgt 240actatcagtg gaagaaatgc ttccggcagt tgcccaagtg ct 282535282DNAZea mays 535caggactgct cattccgggg cctactggct tcaccagacg gatctaaagt atttgagacg 60gcaagaagtg gaccgtactc tttcgacgac atggtcgaga tgggcaaaga cgctggccac 120gaactgaagg cgaaggctgg gcctggcttc ttcgatagcc ttcaatgaac agaatgtgcg 180gccatgcgcg atttcagttg gcaccctttc ggttgaaaac gagggccata gtaggttgtt 240gaggggtttg tttttgtttc ttcttttttt ctcctactac ta 282536174DNAZea mays 536cgggaactgc tcattccggg gcctactgtc ttcaccagac ggatctaaag tatttgagac 60ggcaagaagt ggaccgtact ctttcgacga catggtcgag atgggcaaag acgctggcca 120cgagctgaag gcgaaggctg ggcctggctt cttcgatagc cttcaatgaa caga 174537315DNAZea mays 537cgggaactgc tcattccggg gcctactgtc ttcaccagac ggatctaaag tatttgagac 60ggcaagaagt ggaccgtact ctttcgacga catggtcgag atgggcaaag acgctggcca 120cgagctgaag gcgaaggctg ggcctggctt cttcgatagc cttcaatgaa cagaatgtgc 180ggccatgcgc gatttcagtt ggcacccttt cggttgaaaa cgagggccaa agtaggttgt 240tcaggggctt gtttgtgata cttctgagtt tctcctacta ctaggtcctg ctagagcctt 300gtactaccac tcatg 315538338DNAZea mays 538ctctatgaaa gatgttccaa catatctacc tgaaggcaca atattgccct gtgagctccg 60acgagaagat gtaagagatg cattcatatg cttgactgca aattcgctcg cggagcttcc 120tgctggcagt gttgttggaa gtgcttcctt gcggagacaa tctcagattc tctacagata 180tccatcactg aaagtagtta acttcagagg aaatgttcag acacggttaa agaaactcaa 240ggaaagagat gtgtctgcta cattgttggc gctggctgga ttaaagcggc taaaaatggc 300agaaaatgca acagctgtac tatcagtgga agaaatgc 338539422DNAZea mays 539ccaaggtctc actcatccgg attgggacgc gtgggagtcc tctggctctt gcacaagccg 60atgaaactcg ggaaaaactg aaagccgcac actctgagtt agctgaggag ggggctattg 120agatcgtcat cataaagacc acaggagaca tgatcttgga caaacccctt gcagatattg 180gaggcaaggg tttattcacc aaggagatag atgatgcact cttgcaggga aggattgata 240tagctgtgca ctctatgaaa gatgttccaa catatctacc tgaaggcaca atattgccct 300gtaacctccc acgagaagat gtaagagatg cattcatatg cttgactgca aattcgctcg 360cggagcttcc tgctggcagt gttgttggaa gtgcttcctt gcggagacaa tctcagattc 420tc 422540280DNAZea mays 540ctctggctct tgcacaagcc catgaaactc gggaaaaact gaaagccgca cactctgagt 60tagctgagga gggggctatt gagatcgtca tcataaagac cacaggagac atgatcttgg 120acaaacccct tgcagatatt ggaggcaagg gtttattcac caaggagata gatgatgcac 180tcttgcaggg aaggattgat atagctgtgc actctatgaa agatgttcca acatatctac 240ctgaaggcac aatattgccc tgtaacctcc cacgagaaga 280541255DNAZea maysunsure at all n locations 541gggtttattc accaaggaga tagatgatgc actcttgcag ggaaggattg atatagctgt 60gcactctatg aaagatgttc caacatatct acctgaaggc acaatattgc cctgtaacct 120cccacgagaa gatgtaagag atgcattcat atgcttgact gcaaattcgc tcgcggantt 180cctgctggca gtgttgttgg aagtgcttcc ttgcggagac aatctcagat tctctacaga 240tatccatcac tgaaa 255542269DNAZea mays 542gcactcttgc agggaaggaa tgatatagct gagcactcta tgaaagatgt tccaacataa 60ctacctgaag gcacaatatt gccctgtaac ctcccacgag aagatgtaag agatgcattc 120atatgcttga ctgcaaattc gctcgcggag cttcctgctg gcagtgttgt tggaagtgct 180tccttgcgga gacaatctca gattctctac agatatccat cactgaaagt agttaacttc 240agaggaaatg ttcagacacg gttaaggaa 269543334DNAZea mays 543agagccacgc gtccgcccac gcgtccgcct tgtcaaagcc ggcaatggtg ttgccaccct 60tggcctccct gactcccctg gcttccccaa cggggccacg taccacactt tgacggcacc 120ctacaatgat gtgcaccgca gtgatcaaac tgttcgaaga caaacccgtg gagattgcgg 180gcgtcctcct cgaaccagtt gttggcaacg ctcgtttcat ccctccagag acatggtttc 240cttaacgctc tccgcgactt gaccaggcag gatggtgcgc tccagggcgt cgatgaactg 300atgaccggct tccgtctgtc ttacggtgga cctc 334544429DNAZea maysunsure at all n locations 544cccacgcgtt cggcgggaac cctctagcca tgaccgctgg gatccacacg ctcaagcggc 60tgacagagcc cggcacctac gagtacttgg acaagatcac cggcgaactc gtccgtggga 120tactggacgt cggtgcgaaa gcagggcatg atatgtgcgg aggacatatc agaggaatgt 180ttggcttctt cttcaccggc gggcccgtcc acaacttcgg ggacgccaag aagagcgaca 240ccgagaagtt cgggaggttc taccgtggca tgctggagga gggcgtgtac ttcgctccat 300cgcagttcga ggcggngttc accagcttgg cgcacacctt ccaggacatc gagaagaccg 360tcgaggccgc tgagaaggtt ctgaagcgga tatagggggt ccgcttcaag caagcatgca 420gagagcatt 429545403DNAZea maysunsure at all n locations 545aatgggatcc acacgctcaa gcggctgaca gagcccggca cctacgagta cttggacaag 60atcaccggcg aactcgtccg tgggatactg gacgtcggtg cgaaagcagg gcatgagatg 120tgcggaggac atatcagagg aatgtttggc ttcttcttca ccggcgggcc cgtccacaac 180ttcggggacg ccaagaagag cgacaccgag aagttcggga ggttctaccg tggcatgctg 240gaggagggcg tgtacttcgc tccctcgcag ttcgaggcgg ggttcaccag cttggcgcac 300acctcccagg acatcgagaa gaccgtcgag gccgctgaga aggttctgaa gcggatatan 360ggggtccgct tcaagcaagc atgcagagag catttcctcg tat 403546312DNAZea mays 546agaaactgtt cgaggacaac gcgggggaga ttgctgccgt cttcctcgag ccagttgttg 60gcaacgctgg tttcatcccc ccacagcctg gtttccttaa cgctctccgc gacttgacca 120aacaggatgg tgcgctcctg gtcttcgatg aagtgatgac cggcttccgt ctgtcttacg 180gtggagctca ggagtacttc gggatcaccc ctgacgtgac gaccttgggc aagatcatcg 240ggggtggcct ccccgttggt gcctacggtg ggagaaggga catcatggag atggttgccc 300ccgaaggccg at 312547286DNAZea mays 547ggttgccccc gcaggccgat gtaccaggca ggaactctca gcgggaaccc tctagccatg 60accgctggga tccacacgct caagcggctg acagagcccg gcacctacga gtacttggac 120aagatcaccg gcgaactcgt ccgtgggata ctggacgtcg gtgcgaaagc agggcatgag 180atgtgcggag gacatatcag aggaatgttt ggcttcttct tcaccggcgg gcccgtccac 240aacttcgggg acgccaagaa gagcgacacc gagaagttcg ggaggt 286548285DNAZea mays 548cctgacgtga cgaccttggg caagatcatc gggggtggcc tccccgttgg tgcctacggt 60gggagaaggg acatcatgga gatggttgcc cccgcaggcc gatgtaccag gcaggaactc 120tcagcgggaa ccctctagcc atgaccgctg ggatccacac gctcaagcgg ctgacagagc 180ccggcaccta cgagtacttg gacaagatca ccggcgaact cgtccgtggg atactggacg 240tcggtgcgaa agcagggcat gagatgtgcg gaggacatat cagag 285549243DNAZea mays 549gaccggcttc cgtctgtctt acggtggagc tcaggagtac ttcgggatca cccctgacgt 60gacgaccttg ggcaagatca tcgggggtgg cctccccgtt ggtgcctacg gtgggagaag 120ggacatcatg gagatggttg cccccgcagc cgatgtacca ggcaggaact ctcagcggga 180accctctagc catgaccgct gggatccaca cgctcaagcg gctgacagag cccggcacct 240acg 243550247DNAZea mays 550gtttccttaa cgctctccgc gacttgacca aacaggatgg tgcgctcctg gtcttcgatg 60aagtgatgac cggcttccgt ctgtcttacg gtggagctca ggagtacttc gggatcaccc 120ctgacgtgac gaccttgggc aagatcatcg ggggtggcct ccccgttggt gcctacggtg 180ggagaaggga catcatggag atggttgccc ccgcaggccg atgtaccagg caggaactct 240cagcggg 247551223DNAZea mays 551gcacgaggca gggccgatgt accaggcagg aactctcagc gggaaccctc tagccatgac 60cgctgggatc cacacgctca agcggctgac agagcccggc acctacgagt acttggacaa 120gatcaccggc gaactcgtcc gtgggatact ggacgtcggt gcgaaacagg gcatgagatg 180tgcggaggac atatcagagg aatgtttggc ttcttcttca ccg 223552218DNAZea mays 552gcacgaggca gggccgatgt accaggcagg aactctcagc gggaaccctc tagccatgac 60cgctgggatc cacacgctca agcggctgac agagcccggc acctacgagt acttggacaa 120gatcaccggc gaactcgtcc gtgggatact ggacgtcggt gcgaaagcag ggcatgagat 180gtgcggagga catatcagag gaatgtttgg cttcttct 218553275DNAZea mays 553gcgaaacagg gcatgagatg tgcggaggac atatcagagg aatgtttggc ttctacttca 60ccggcgggcc cgtccacaac ttcggggacg ccaagaagag cgacaccgag aagttacaga 120ggttctaccg tggcatgctg gaagaggcgt gtacttcgct ccctcgcagt tcgaggcggg 180gttcaccagc ttggcgcaca cctcccagga catcgagaag accgtcgagg ccgtaatgaa 240ggttctgaag cggatatagg gggtacgctt caagc 275554252DNAZea mays

554cttcggggac gccaagaaga gcgacaccga gaagttcggg aggttctacc gtggcatgct 60ggaggagggc gtgtacttcg ctccctcgca gttcgaggcg gggttcacca gcttggcgca 120cacctcccag gacatcgaga agaccgtcga ggccgctgag aaggttctga agcggatata 180gggggtccgc ttcaagcaag catgcagaga gcatttcctc gtatctacgt tcttgtactc 240ttagttctat at 252555295DNAZea mays 555ctctagccat gaccgctggg atccacacgc tcaagcggct gacagagccc ggcacctacg 60agtacttgga caagatcacc ggcgaactcg tccgtgggat actggacgtc ggtgcgaaag 120cagggcatga gatgtgcgga ggacatatca gaggaatgtt tggcttcttc ttcaccggcg 180ggcccgtcca caacttcggg gacgccaaga agagcgacac cgagaagttc gggaggttct 240acgtggcatg cctggagagg gcgtgtactt cggctccctc gcagttcgag gcggg 295556331DNAZea mays 556ccacgcgtcc gagggcgtgt acttcgctcc ctcgcagttc gaggcggggt tcaccagctt 60ggcgcacacc tcccaggaca tcgagaagac cgtcgaggca gctgagaagg ttctgaagcg 120gatatagggg gtccgcttca agcaagcatg cagagagcat ttcctcgtat ctacgttctt 180gtactcttag ttctatatgc caccgaggtt ttgtattgtg cagcagcagg acagcttctg 240taagttcctc tttctgaatt agtgggtctt gtttttgtca gtgccaataa atctctggtc 300cacgattacg gtttcgttgt tgtactgatg t 331557423DNAZea mays 557gacccaatcg ccgcaaaccc ctccggaatt tcttatcccc cctcatctgc tccacctccg 60acctcgcgcg agacgagcaa gcccaagtat ggccggagca gcagcagccg ccgtggcgtc 120cggggtctcg gcccggccgg ccgcgccgag gagggcttct gcgggacgcc gcgctcggct 180gtcggtggtg cgggccgcga tatccctcga gaagggcgag aaggcgtaca cggtgcagaa 240gtccgaggag atcttcaacg ccgccaagga gctgatgcct ggaggtgtta actcgccagt 300ccgagccttc aaatctgttg gtgggcagcc agtagttttc gactctgtaa agggttctcg 360tatgtgggat gttgatggga atgagtacat tgattacgtt ggttcctggg gtcctgcaat 420cat 423558302DNAZea mays 558cggacgcgtg ggcggacgcg tgggcgccga ggagggcttc tgcgggacgc cgcgctcggc 60tgtcggtggt gcgggccgcg atatccctcg agaagggcga gatagcgtac acggtgcagc 120agtccgagga gatcttcaac gccgccaatg agctgatgcc tggaggtgtt aactcgccag 180tccgagcctt caaatctgtt ggtgggcagc cagtagtttt cgactctgta aagggttctc 240gtatgtggga tgttgatggg aatgagtaca ttgattacgt tggttcctgg ggtcctgcaa 300tc 302559305DNAZea maysunsure at all n locations 559ctgctccacc tccgacctcg cgcgagacga gcaagcccaa gtatggccgg agcagcagca 60gccgccgtgg cgtccggagt ctcggcccgg ccggccgcgc cgaggagggc ttctgcggga 120cgccgcgctc ggctgtcggt ggtgcgggcc gcgatatccc tcgagaangg cgagaaggcg 180tacacggtgc agaagtccga ggagatcttc aaggccgcca aggagctgat gcctggaggt 240gttaactcgc cagtccgagg cttcaaatct gttggtgggc agccagtagt ttcgactctg 300taaag 305560276DNAZea mays 560gctccacctc cgacctcgcg cgagacgagc aagcccaagt atggccggag cagcagcagc 60cgccgtggcg tccggggtct cggcccggcc ggccgcgccg aggagggctt ctgcgggacg 120ccgcgctcgg ctgtcggtgg tgcgggccgc gatatccctc gagaagggcg agaaggcgta 180cacggtgcag aagtccgagg agatcttcaa cgccgccaag gagctgatgc ctggaggtgt 240taactcgcca gtccgagcct tcaaatctgt tggtgg 276561225DNAZea mays 561cccacgcgtc cgcccacgcg tccgcccacg cgtccgctgc gggacccgcg ctcggctgtc 60ggtggtgcgg gccgcgatat ccctcgagaa gggcgagaag gcgtacacgg tgcagaagtc 120cgaggagatc ttcaacgccg ccaaggagct gatgcctgga ggtgttaact cgccagtccg 180agccttcaaa tctgtatgtg ggcagccagt agttttcgac tctgt 225562276DNAZea mays 562cagacgcgtg ggcgagacgc gtgggctgct ccacctccga cctcgcgcga gacgagcaag 60cccaagtatg gccggagcag cagcagccgc cgtggcgtcc ggggtctaca cccggccgga 120cgcgccgagg agggcttctg cgggacgccg cgctcggctg tcggtggtgc gggccgcgat 180atccctcgag aagggcgaga aggcgtacac ggtgcagaag tccgaggaga tcttcaacgc 240cgccaaggag ctgatgcctg gaggtgttaa ctcgcc 276563251DNAZea mays 563ccacgcgtcc gtccacctcc gacctcgcgc gagacgagca agcccaagta tggccggagc 60agcagcagcc gccgtggcgt ccggggtctc ggcccggccg gccgcgccga ggagggcttc 120tgcgggacgc cgcgctcggc tgtcggtggt gcgggccgcg atatccctcg agaagggcga 180gaaggcgtac acggtgcaga agtccgagga gatcttcaac gccgccaagg agctgatgcc 240tggaggtgtt a 251564337DNAZea mays 564caagtatcga aatggtccgc tttgtcaact cagggacaga agcctgcatg ggagcgctcc 60gcctcgtgcg cgcattcacc gggcgggaga agatcatcaa gttcgaaggc tgctaccatg 120gccatgccga ttccttcctt gtcaaagccg gcagtggtgt tgccaccctt ggcatcactg 180actcccctgg cgtccccaag ggggccacct acgagacttt gacggcaccc tacaatgatg 240tcgcggcagt gaagaaactg ttcgacgaca acgcggggga gattgctgcc gtcttcctcg 300agtcagttgt tggcaacgct ggtttcaatc ccccaca 337565263DNAZea mays 565gaaactctga agaaaggaac tagctttggt gctccatgtt tgctggagaa cgtattggct 60gagatggtca tctctgccgt gccaagtatc gaaatggtcc gctttgtcaa ctcagggaca 120gaagcctgca tgggagcgct ccgcctcgtg cgcgcattca ccgggcggga gaagatcatc 180aagttcgaag gctgctacca tggccatgcc gattccttcc ttgtcaaagc cggcagtggt 240gttgccaccc ttggcctccc tga 263566310DNAZea mays 566gaacaccacg aatcgtctgc attcggctcg aggacactct gaagaaagga actagctttg 60gtgctccatg tttgctggag aacgtattgg ctgagatggt catctctgcc gtgccaagta 120tcgaaatggt ccgctttgtc aactcaggga cagaagcctg catgggagcg ctccgcctcg 180tgcgcgcatt caccgggcgg gagaagatca tcaagttcga aggctgctac catggccatg 240ccgattcctt ccttgtcaaa gccggcagtg gtgttgccac ccttggcctc cctgactccc 300ctggcgtccc 310567124DNAZea mays 567gctttgtcaa ctcagggaca gaagcctgca tgggagcgct ccgcctcgtg cgcgcattca 60ccgggcggga gaagatcatc aagttcgaag gctgctacca tggccatggc gaatccttcc 120ttgt 124568295DNAZea maysunsure at all n locations 568cggacgcgtg gcgagacgcg tgggcggacg cgtgggcctt gtcaaagccg gcagtggtgt 60tgccaccctt ggcctccctg actcccctgg cgtcccacac ggggccacca cctgagactt 120tgacangaac cctacaatga tgtcgcggca gtgaagaaac tgttcgagga caacgcgggg 180gagattgctg ccgtcttcct cgagccagtt gttggcaacg ctggtttcat ccccccacag 240cctggtttcc ttaacgctct ccgcgacttg accaaacagg atggtgcgct cctgg 295569253DNAZea mays 569cccacgcgtc cgcccacgcg tccgctcccc tggcgtcccc aagggggcca cctacgagac 60tttgacggca ccctacaatg atgtcgcggc agtgaagaaa ctgttcgagg acaacgcggg 120ggagattgct gccgtcttcc tcgagccagt tgttggcaac gctggtttca tccccccaca 180gcctggtttc cttaacgctc tccgcgactt gaccaaacag gatggtgcgc tcctggtctt 240cgatgaagtg atg 253570363DNAZea mays 570ggtgcacggt agtgagtcgg aatcggctcg agtggcgatg gaaatctggg agctactgaa 60agaattcttt gatgcagaaa ttagaaagct gaagctacaa ccatattatt tcgctattgt 120tgttactgag aatgttctac agaaggaaaa ggaccacatt gagggctttg cacctgaggt 180agcttgggtt actaaatctg ggaaatctga cctggaagca ccgattgcaa gtgcgcccac 240aggtgagctt gtaatgaacc cggctttctc catatggata agacgccacc gagacttacc 300cttgaggtgt aatcaatggt gtcatgttgt tagatgggag tttagcgatc cgactccttt 360cat 363571312DNAZea mays 571accacgcgtc cgcccacgcg tccgagaagc aggaattaga gttaaagtgg acgactcaga 60gctgcgaact cctggatgga aattcaatca ctatgagatg aaaggggttc ctgtaagaat 120atagataggt ccacgtgatg tcacaaataa gagtgttgtg gtttctaggc gtgatgtccc 180tggaaagcaa ggaaaggagt ttggagtgtc tatggagcct tcgatattgg tgaaccatat 240aaatggtcgt ctagatgaca tacaagcatg ccttttacag aaggccttaa aatccgtgat 300agtaacattg tc 312572270DNAZea maysunsure at all n locations 572ttaacttgca nngccaggtc aaggtctaga attcccaggc cgacctacga ctacacgtcg 60gcccacccgt ccggccaaga tggctcctga gggctaagaa aagctgtaca ccaaggtcaa 120gagcattcac gacagcctga tcgaggctgg tgtccgcgtc gagtccgact accgtgaggg 180ctactccccc ggatggaagt tcaacgactg ggagctcaag ggtaatcctc ttcctaacca 240attccgtccc aaggattccc aaaaaggttt 270573427DNAZea mays 573cccacgcgtc cgcccacgcg tccgcccacg cgtccgccca cgcgtccgtg ggaaaatgtg 60gccagatgct tctgatactg atgcttcctc tcactataag cttccgttct caagaactgt 120ctacattgag aaaactgatt ttcgccttaa ggactcaaaa gactactatg ggctggcccc 180tggtaaatct gtcatgctaa ggtatgcgtt ccccataaaa tgcacagacg ttatctatgg 240tgatactcct gatgatattg ttgaaattcg agcagaatat gatcctttga agacttctaa 300acttaagggt gttctgcact gggttgctga gccagcacct ggtgtcgaac cattgaaggt 360ggaagtaaga ctattcgaga aattgttcat gtcagagaat cctgctgaat tggaggattg 420gcttggt 427574273DNAZea mays 574gttgaggaga gtggaaattt atgaattcag ccgattgaat atggtttaca ctcttctaag 60caagcgaaag cttctttggt ttgtacaaaa caagaaggtc gaagattgga cagacccacg 120ttttcccact gtccaaggca tagtacgtcg gggcttgaag gttgatgcat tgatacagtt 180tatactccaa cagggtgctt caaaaaatct gaatctcatg gaatgggata aactctggac 240aatcaacaag aagataattg atccagtgtg cgc 273575267DNAZea mays 575cccacgcgtc cggacggtat tgagtcaagg tgcagaaata ataccgtgga ggaaaatctc 60tcattatgga aagagatggt taatggaact gaaaggggca tgcagtgctg tgtacggggt 120aaacttgaca tgcaggatcc taacaagtca ctcagggatc ctgtttacta ccgctgtaat 180actgatccac accatcgtgt tggttcgaag tacaaggtct atccaacata tgactttgcg 240tgcccatttg tcgatgcatt ggagggg 267576380DNAZea mays 576cggacgcgtg ggctgctgaa ttggaagatt ggcttggcga tcttaaccca cactcgaaag 60aggtgataaa ggatgcttat gctgtaccat cacttgccac tgcggttctg ggtgacaagt 120tccagtttga gcggcttggt tacttcgccg tggatactga ctccacacct gagaaactcg 180tgttcaacag aactgttacc ctccgtgatt cgttcgggaa agctggaccc aagtgactgt 240tcagtgtaat ttagggaggg cgctggtttt gatcggttgc agaagcgcac ctgaactata 300caagttgtga agaaaatggt cgtctaatac agaacagttt aaagggcctt actctttata 360aaatttaggg ttttttaaaa 380577373DNAZea mays 577actgtttaca cactcaatca atctgggatt tgagcggatc aggacacccg tgaaaattag 60ctctccaggt tggaagtatt ctcactggga aatgaaaggt gttccattga gaattgagat 120tggtccaaaa gatctggcaa acaaacaggt acgcattgtc cgccgggaca acggtgcaaa 180ggttgacatt ccggtgacca atttggttga agatgttaaa gtgttattgg atgagattca 240aaaaaatctg ttcaaaacag ctcaagaaag gagagatgca tgtgttcagg tcgtcaactc 300ttgggatgaa ttcacaactg ctctgaataa caaaaggttg atcttggctc cttggtgcga 360tgaggaggaa gtt 373578299DNAZea mays 578cgtgattcca gtgccttata aggacgctga cacaactgcc ataaagggag cctgcgaatc 60aactgtttac acactcaatc aatctgggat tcgagcggat caggacaccc gtgaaaatta 120ctctccaggt tggaagtatt ctcactggga aatgaaaggt gttccattga gaattgagat 180tggtccaaaa gatctggcaa acaaacaggt acgcattgtc cgccgggaca acggtgcaaa 240ggttgacatt ccggtgacca atttggttga agatgttaaa gtgttattgg atgagattc 299579286DNAZea mays 579gccaatccag gtaattgtga ttccagtgcc ttataaggat gctgacacaa ctgccataaa 60gggagcctgc gaatcaactg tttacacact cgatcaatct ggaattagag cggatcagga 120cacccgtgaa aattactctc caggttggaa gtattcccac tgggaaatga aaggtgttcc 180attgagaatt gagattggtc caaaagatct ggcaaacaaa caggtgcgtg ttgtccgccg 240ggacaacggt gcaaaggttg acatccctgt gaccaatttg gttgaa 286580313DNAZea mays 580gatgacaaag gcttagtatt accaccaaag gtagcgccaa tccaggtaat tgtgattcca 60gtgccttata aggatgctga cacaactgcc ataaagggag cctgcgaatc aactgtttac 120acactcgatc aatctggaat tagagcggat caggacaccc gtgaaaatta ctctccaggt 180tggaagtatt cccactggga aatgaaaggt gttccattga gaattgagat tggtccaaaa 240gatctggcaa acaaacaggt gcgtgttgtc cgccgggaca acggtgcaaa ggttgacatc 300cctgtgacca att 313581307DNAZea mays 581cccacgcgtc cgcacatggt gatgacaaag gcttagtatt accaccaaag gtagcgccaa 60tccaggtaat tgtgattcca gtgccttata aggatgctga cacaactgcc ataaagggag 120cctgcgaatc aactgtttac acactcgatc aatctggaat tagagcggat caggacaccc 180gtgaaaatta ctctccaggt tggaagtatt cccactggga aatgaaaggt gttccattga 240gaattgagat tggtccaaaa gatctggcaa acaaacaggt gcgtgttgtc cgccgggaca 300acggtgc 307582227DNAZea mays 582cccacgcgtc cggaaaggtg ttccattgag aattgagatt ggtccaaaag atctggcaaa 60caaacaggtg cgtgttgtcc gccgggacaa cggtgcaaag gttgacatcc ctgtgaccaa 120tttggttgaa gaggttaaag tgttactgga tgagattcaa aaaaatctgt tcaaaacagc 180ccaagaaaag agagatgcct gtgttcatgt cgtgaacact tgggatg 227583427DNAZea mays 583ggttgacaat attacatgtg caccgaccac aaaccaaata atcagcaaaa tggatttcga 60gtggcatctc aacatgcaca accttaggta aaagcttgag atggagaaac taaaagtttc 120caacagcgaa cacaaagagt ggctggggct ggcctaggag gggaggaaga agagtgccat 180cacacgaaaa ccatgacctc acagcattgg tgcagtaaca tttcactatt tagagcctat 240gatcaggctt taaagagtgg ctggggctgg cctaggaggg gaggaagaag agtgccatca 300ctaacaaaac agcccctcga accatggttg ttttgcgacc tctaaaggtg gtaataacta 360acttggaaga aggaaaagta ctagaccttg atggcaaaat gtggcctgat gcttctgata 420ctgatgc 427584499DNAZea mays 584tgggtagtgt aacatcacaa tgctactgcc aactcatata ctaggactcg ttggtcgtta 60caacactcta gattcactcg tattaaccga atctgtgagc catgtcgacc aacaagggca 120gcgcggccaa gggcggcgga gggaagaaga aggaggtgaa gaaggagacg aagctcggga 180tggcctataa gaaggacgac aacttcgggg agtggtactc cgaggttgtt gttaacagtg 240aaatgattga gtactatgac atttctggtt gttatatatt gaggccatgg gcgatggaaa 300tctgggagct actgaaagaa ttctttgatg cagaaattaa aaagctgaag ctcaaaccat 360attatttccc tttgtttgtt actgagaatg ttctacagaa ggaaaaggac cacattgagg 420gctttgcacc tgaggtagct tgggttacta aatctgggaa atctgacctg gaagcaccga 480ttgcaatccg ccccacaag 499585284DNAZea mays 585gacatttctg gttgttatat attgaggcca tgggcgatgg aaatctggga gctactgaaa 60gaattctttg atgcagaaat taaaaagctg aagctcaaac catattattt ccctttgttt 120gttactgaga atgttctaca gaaggaaaag gaccacattg agggctttgc acctgaggta 180gcttgggtta ctaaatctgg gaaatctgac ctggaagcac cgattgcaat ccgccccaca 240agtgagactg tcatgtatcc gtacttctcc aaatggataa gaag 284586271DNAZea mays 586ggaccgtggc ggtacgcgtg ggtttgtcga catatctgtc ccaaggaatg tcagcgcgtg 60cgtctctgaa attggctccg agcgagtata caatgtcgac gacctgaaag aggtggtgga 120agccaacaag gaagaccgtc tcaggaaagc gatggaggca cagacaatca tcgccgaaga 180gctgaaacgg tttgaggcgt ggcgggactc gctggagacc gttccaacca tcaagaagct 240gaggtcttac gccgacagga tccgggcctc g 271587230DNAZea mays 587accatattga agaggctgct gtgcttagac ctgtaacaga atggaaattt atgtggtggc 60cctatcatgg aaccgaggta tcagggaagt cgtggactgg atgtcgaaga aaagtggtat 120tcctgcttct gagcttaggg aacacctatt catgctgcgt gacagtgatg ctacacgcca 180tctgtttgag gtatcggctg ggttggactc tctggttctc ggtgaaggac 230588229DNAZea mays 588gtggccccgt gctattcaag aactcactag cctgaaccat attgaagagg ctgctgttct 60tagtacctgt aatagaatgg aaatttatgt ggtggcgcta tcatggaacc gtggtatcag 120agaagtagtg gactggatgt cgaagaaaag tggtattccc gcttccgagc ttagggagca 180cctgttcatc ttgcgaacag tgatgccaca cgccatctgt ttgaggtgt 229589492DNAZea maysunsure at all n locations 589aggttaaagt ntgtaataga tgggatgtac tgtacacttc tccggnttnn nnnnnnggng 60gggagccacg cgtccggaaa tgttaacgca ttaaaaggta tacggtatca gtaaacctta 120caagtgtgat gccaagggaa aacggcatca gctgacacat tgctatattc ctgtttattt 180cgtccgaata aagtatataa cttaagaaag gggctcttgc cccacagcag ctcaagcaaa 240aatgtacaaa gaaaagcagc tcgagtagag agaatttgcc actctctcga cagattgagc 300tgctgccatg gcgctaattc acgacacatt tgatgtctcg gcaagacggg gaggagctca 360gtaagtgaga tgataaaaaa atagaatcag gttggagggt aagtatacac gggtagaaaa 420attgcctcct tggccttaat tntgggtctt ctccaccttg gccttgatct tctgctcgat 480gattgccttc tc 492590313DNAZea mays 590cgtggaaaac tttccggttc caaaggacct ttggcccctt ccttttaaga acctacctgg 60gtaaaccctt tttgaaaagg ctcctgtcct aatacttgta taaaatgaaa attatgtggt 120agccctatca tggaaccgaa gtatcagaga agtagttgac tggatgtcaa agaaaagtgg 180tattcctgct tctgagctta aggagcacct attcatgctg cgtgacagtg atgctacacg 240ccatctgttc taagtatcag caaggttgga ctctttggtt ctcggtgaac gacaaatcct 300tgctcaagtc aaa 313591457DNAZea mays 591agcccacgcg tccgcccacg cgtccggtga aatcccgcac ctacctcctt cctctctcac 60cgaggaccct cgcaccaaga actgagcggg aagagaggta gagaggcaag cgcacgagag 120tttctgctcc tagtctcgtc tcgccccgcc tccgtctcct ttccctctct ggttctctct 180ctgcgattct cgtcgcattg gttccgttcc ctcacgaaag gcggtagctt tctgtcttcc 240ctgatctatc tagataatgg cgaccacgac gtcagcgacc accgccgcag cagcagccgc 300caccatcgcc aagccgcggg ggtcgtcgtc ggacctctgc cagagggtgg ccggcggcgg 360caggcggtgc tccggggtgg tgccgtgcga cgccgccggc gtggaggccc aggcgcatgc 420cgtggcaaat gcggccagcg tcgccgccct cgagcag 457592267DNAZea mays 592gaaggttgtt gtggtgaacc gctccgtgga aagggtggat gctattcgtg aggagatgaa 60agatatagag atcgtgtaca ggcctctctc agacatgtat caagctgctg ctgaagctga 120tgtcgtgttc accagcaccg catctgaaac ttcattgttc gcaaaagaac acgcagaggc 180actcccccct gtctctgata ctatgggagg tgttcgcctg tttgtcgaca tatctgtccc 240caggaatgtc agcgcatgtg tgtctga 267593264DNAZea mays 593cccacgcgtc cgcccacgcg tccgggatgc aagaaggttg ttgtggtgaa ccgctccgtg 60gaaagggtgg atgctattcg tgaggagatg aaagatatag agatcgtgta caggcctctc 120tcagacatgt atcaagctgc tgctgaagct gatgtcgtgt tcaccagcac cgcatctgaa 180acttcattgt tcgcaaaaga acacgcagag gcactccccc ctgtctctga tactatggga 240ggtgttcgcc tgtttgtcga cata 264594310DNAZea mays 594atcttattgc caaaggatgc aagaaggtgg ttgtggtcaa ccgttcagtg gaaagggtgg 60atgccatccg cgaggagatg aaaggtatcg agattgtgta caggcctctt tcagagatgt 120acgaagctgc tgctgaagct gatgtcctat tcacgagcac tgcatctgaa accccattgt 180tcacaaaaga gcacgcagag gcacttccca caatttccga tgccatggat ggtgcccggc 240tttttgtcga catatctgtc ccaaggaatg tcagcgcgtg cgtctctgaa attggctccg 300cgcgagtata 310595290DNAZea mays 595gtggtcaacc gttcagcaca aagggtggat gccatccgcg aggagattaa agctatcgag

60attgtgtaca ggcctctctc ggagatgtat gaagctgctg ctgaagctga cgtcgtgttc 120acgagcaccg catctgaaac cccattgttc acaaaagagc acgcagatgc acttcccact 180gtttctgatg ccatgggcgg tgtccggctc tttgtcgaca tatctgtccc aaggaatgtc 240agcgcgtgtg tctctgaaat tggctccgcg cgagtgtaca atgttgatga 290596168DNAZea mays 596ggtggttgtg gtcaaccgtt cagtggaaag ggtggatgcc atccgcgagg agatgaaagg 60tatcgagatt gtgtacaggc ctctttcaga gatgtacgaa gctgctgctg aagctgatgt 120cctattcacg agcactgcat ctgaaacccc attgttcaca aaagagca 168597254DNAZea mays 597acctgaaaga ggtggtggaa gccaacaagg aagaccgtct caggaaagcg atggaggcac 60agacaatcat cgccgaagag ctgaaacggt ttgaggcgtg gcgggactcg ctggagaccg 120ttccaaccat caagaagctg aggtcttacg ccgacaggat ccgggcctcg gagctcgaga 180agtgcctgca gaagatcggg gacgacgctc tcaccaagaa gacgaggaga gccatcgagg 240agctaagcac cggc 254598270DNAZea mays 598cggctcgagg aaagaggtgg tggaagccaa caaggaagac cgtctcagga aggcaatgga 60ggcgcagaca atcatcaccg aagagctgaa acggtttgag gcatggcggg actcgctgga 120gaccgttcca accatcaaga agctgaggtc atatgccgac aggatccgag cctcagagct 180cgatgagtgc ctacagaaga tcggggatga cgttctcacc aagaagatga ggagagccat 240cgaggagcta agcaccggca tcgtgaacaa 270599422DNAZea mays 599cgaccatcaa gaagctgagg tcgtacgcgg acaggatcag ggcctcggag ctcgagaagt 60gcctgcagaa agtaggtgag gacgccctca ccaagaagat gaggagagcc atcgaggagc 120tgagcaccgg catcgttaac aagctcctcc atggcccgct gcagcacctg aggtgcgacg 180gcagcgacag ccgcaccctt gacgagacgc tcgagaacat gcacgccctc aaccggatgt 240tcagcctcga catggagaag gcgatcatcg agcagaagat caaggccaag gtggagaaga 300cacaaaactg aggccaggaa gcaatttttc taccaccatt atctatatat atagcgtctc 360caatctcatt ccattttttt atcctttcac tcagtgagcc cttcccctgc tcactgtgat 420cg 422600282DNAZea mays 600gacaggatca gggcctcgga gctcgagaag tgcctgcaga aagtaggtga ggacgccctc 60accaagaaga tgaggagagc catcgaggag ctgagcaccg gcatcgttaa caagctcctc 120catggcccgc tgcagcacct gaggtgcgac ggcagcgaca gccgcaccct tgacgagacg 180ctcgagaaca tgcacgctct caaccggatg ttcagcctcg acatggagaa ggcgatcatc 240gagcagaaga tcaaggccaa ggtggagaag acacaaaact ga 282601262DNAZea mays 601tgacgttctc accaagaaga tgaggagagc catcgaggag ctaagcaccg gcatcgtgaa 60caagctcctc cacggcccgc tgcagcacct gaggtgcgac ggtagtaaca gccgcaccct 120tgatgagacg ctcgagaaca tgcatgctct caaccggatg ttcagcctcg acacggagaa 180ggcgatcatc gagcagaaga tcaaggccaa ggtggagaag acccagaatt gaggcctgga 240gtcaattttt ctacccgtgt at 262602288DNAZea mays 602gacgccctca ccaagaagat gaggagagcc atcgaggagc tgagcaccgg catcgttaac 60aagctcctcc atggcccgct gcagcacctg atgctggacg gcagcgacag ccgcaccctt 120gacgagacgc tcgagaacat gcacgccctc aaccggatgt tcagcctcga catggagaag 180gcgatcatcg agcagaagat caaggccaag gtggagaaga cacaaaactg aggccaggaa 240gcaatttttc taccaccatt atctatatat atagcgtctc caatctca 288603139DNAZea mays 603cgatcatcga gcagaagatc aaggccaagg tggagaagac acaaaactga ggccaggaag 60caatttttct accaccatta tctatatata tagcgtctcc aatctcattc cattttttta 120tcctttcact cagtgagcc 139604460DNAZea mays 604cccacgcgtg cgcccactcg tccggtggta ttcccgcttg cgagcttagg gagcacctgg 60tcatcttgcg aagcagtgat gccacacgcc atctgtttga ggtgtcagct ggccttgact 120ctttggttct cggtgaagga caaatccttg ctcaggttaa acaagttgtg aggagtggac 180agaacagtgg aggcttggga aagaacattg ataggatgtt caaggatgca atcactgctg 240gaaagcgtgt ccgctgcgag accaacatat catctggtgc tgtttctgtc agttcagcgg 300cggttgaact ggccctgatg aagcttccga agtctgaagc actgtcagct aggatgcttc 360tgattggtgc tggtaaaatg ggaaagctag tgatcaaaca tctggttgcc aaaggatgca 420tgaaggttgt tgtggtgaac cgctccgtgg aaagggtgga 460605322DNAZea mays 605aacaagttgt gaggagtgga cagaacagtg gaggcttggg aaagaacatc gataggatgt 60tcaaggatgc aatcactgct ggaaagcgtg tccgcagcga gaccaacata tcatctggtg 120ctgtttctgt cagttcagcg gcggttgaac tggccctgat gaagcttccg aagtctgaag 180cactgtcagc taggatgctt ctgattggtg ctggtaaaat gggaaagcta gtgatcaaac 240atctggttgc caaaggatgc aagaaggttg ttgtggtgaa ccgctccgtg gaaagggtgg 300atgctattcg tgaggagatg aa 322606310DNAZea mays 606tcccgcttcc gagcttaggg agcacctgtt catcttgcga agcagtgatg ccacacgcca 60tctgtttgag gtgtcagctg gccttgactc tttggttctc ggtgaaggac aaatccttgc 120tcaggttaaa caagttgtga ggagtggaca gaacagtgga ggcttgggaa agaacattga 180taggatgttc aaggatgcaa tcactgctgg aaagcgtgtc cgctgcgaga ccaacatatc 240atctggtgct gtttctgtca gttcagcggc ggttgaactg gccctgatga agcttccgaa 300gtctgaagca 310607298DNAZea mays 607gtgaaggaca aatccttgct caggttaaac aagttgtgag gagtggacag aacagtggag 60gcttgggaaa gaacatcgat aggatgttca aggatgcaat cactgctgga aagcgtgtcc 120gcagcgagac caacatatca tctggtgctg tttctgtcag ttcagcggcg gttgaactgg 180ccctgatgaa gcttccgaag tctgaagcac tgtcagctag gatgcttctg attggtgctg 240gtaaaatggg aaagctagtg atcaaacatc tggttgccaa aggatgcaag aaggttgt 298608300DNAZea mays 608agcgtgtccg cagcgagacc aacatatcat ctggtgctgt ttctgtcagt tcagcggcgg 60ttgaactggc cctgatgaag cttccgaagt ctgaagcact gtcagctagg atgcttctga 120ttggtgctgg taaaatggga aagctagtga tcaaacatct ggttgcgaaa ggatgcaaga 180aggttgttgt ggtgaaccgc tccgtggaaa gggtggatgc tattcgtgag gagatgaaag 240atatagagat cgtgtacagg cctctctcag acatgtatca agctgctgct gaagctgatg 300609234DNAZea mays 609gttgaactgg ccctgatgaa gcttccgaag tctgaagcac tgtcagctag gatgcttctg 60attggtgctg gtaaaatggg aaagctagtg atcaaacatc tggttgccaa aggatgcaag 120aaggttgttg tggtgaaccg ctccgtggaa agggtggatg ctattcgtga ggagatgaaa 180gatatagaga tcgtgtacag gcctctctca gacatgtatc aagctgctgc tgaa 234610278DNAZea mays 610cgtgagactg gcggtggata acgcgtcatg gaccgacgat aagcagctcc aggacatgta 60cctgatctgc aagtccgtcg cgatgcgaca tcgacgcacc tgggagcggg catgagagga 120gaagctcaag gcgttcgagc tcgcactggc gacggcagac gccacgttct agaacctcga 180ctcgtcggag atctcactga cggacgtgag ccactacttc gactcggacc cgatcaagct 240cgtgcattgg ctgctcaaag acgggcgagc ggcgtcct 278611251DNAZea mays 611gaagatgtgt acaggggaag tgacaagggc atactggctg acgtcgagct tctgaggcag 60atcactgagg cttcgcgcgg cgccatcacc gccttcgttg agaagaccac aaacagcaaa 120gggcaagtcg tcaatgttac caacaacctc agcaagatac ttggtttcgg tctgtcggaa 180ccatgggtgc agtacctgtc cacgaccaag ttcgtcagag cggacagaga gaagatgagg 240gttctgtttg g 251612126DNAZea mays 612gttctagatc gccagtctct tctcctcctt agttttcctc ttcagttctg cccatctgat 60ggctctagtg cagagctgct ccactctctt gtgcaatgca tgtgacttcc ctgtcctggg 120gtcccg 126613296DNAZea mays 613acgggatttg ccaaggatac aaacttgttc tcagtgtcga tgacaagaag ggacattcct 60gccttgtcat cgaactgaga caagtgtatc cacgggattt gccaaggaaa ttgcaagggt 120tgcccagggg aaatattatt acctccctaa tgcttcagat gctgtaattt ctgctgactc 180caagaccgcc ctgacagact tgaagagctc atgattttgc agcagcggca cccgttttct 240gtaccttttg atagggatgg tgaaccttca ttcatgcagt aatttttgcg taggcc 296614286DNAZea mays 614gtgaacactt gcttgatcgt attgcaatta atttaagtgc tgatcttcca atgagttttg 60atgaccgcgt tgaagcagtg gatattgcaa cacggtttca ggagtctagc aaagaagttt 120tcaaattggt ggaagaaaaa actgaaactg caaaaactca gataattttt gcaagagagt 180atctgaagga tgttactatt agcacagagc agctcaaata tcttgtcatg gaagctatac 240gaggtggctg tcaggggcat cgtgctgagt tgtatgctgc ccgagt 286615239DNAZea mays 615cggacgcgtg gcaaccacgg ctgccttgaa gagcgccaag atcgtcgtgg accgtctcct 60ggagaggcag acggctgaca atggcggcaa gtaccctgag acggtcgcac ttgtcctgtg 120gggcaccgac aacatcaaga cctatggtga gtcactagcc caggtgctgt ggatgattgg 180agttcggcca gttgccgaca ccttcggccg tgtcaaccgt gtggagcctg tcagccttg 239616233DNAZea mays 616gggagtgctt gaagctcgtg gtacaggaca atgagctggg cagcggcaga ggctactggg 60agacatcgga ggagaacctg gacaggctca gggagctcta ctcggaggtt gaagacaaga 120ttgaggggat tgaccggtaa accgatttgc cagattcaaa ggaatgagaa gcttggaact 180cttgtgtctc attgaggctc ttgtacaatg tgtgtgtagc ttatatatat ata 233617302DNAZea maysunsure at all n locations 617cggacgctgc gggtacgaga gggctcgttt cgacagggat ccgaagacgt tccgtgagtc 60gtatcatgac gatcangaga atctccagca gcagatatca tctgcacgga gtaaccttgg 120cgctgtgcag attgaccatg acctccgtgt caagatatcc aaggtgtgct ctgagttgaa 180cgttgatgga ctcagaggtg acattgtgac taacatggct gccaaggcgc tggctgcgtt 240gaaaagaatg gacagcgtca ccgtggagga cattgctact gtcattccca actgcttgag 300gc 302618261DNAZea maysunsure at all n locations 618gtttgggttc ttgggggagt gcctgangct cgtcgtgcaa gacaacgagc tgggaagctt 60gaagcttgcc ctcgagggaa gctacgtcga gcctngccct ngcggcganc cgatncgtan 120cncnaagngc tcccgacagg gnagancatc canntctcga tncgcaggtt atccnaaaca 180aagctncctt tnaagaancc aaaatngnnn gtggncnggt tncttggagn ngtgaaggnt 240ggaanatgng gaaantaccc g 261619262DNAZea mays 619ggggcatcgt gctgagttgt atgctgcccg agttgcaaaa tgtctagctg ctatggaagg 60acgtgaaaaa gtatttgtgg atgacctcaa gaaagctgta gagctggtca ttctacctcg 120ctccatccta tctgataatc cacaggatca gcagcaagag catccacccc cacccccgcc 180gccaccacct ccagaaaatc aagattcttc agaagaccaa gatgaggaag acgaagacca 240agaggatgat gaagaagaaa at 262620125DNAZea maysunsure at all n locations 620ccagttctgg ctcggcggct cgtcggacaa tctccagaac ttccttaaga tgatcggcgg 60ctggtacntg cctgccctca aaggcgccgg catcaagtac gacgaccccc gtgctctacc 120tcgac 125621280DNAZea mays 621gcaagggttg cccaggggaa atattattac ctccctaatg cttcagatgc tgtaatttct 60gctgccacca agaccgccct gacagacttg aagagctcat gattttgcag cagcggcacc 120cgttttctgt accttttgat agggatggtg aaccttcatt catgcagtaa tttttgcgta 180ggcctctaca atgacagggg gaaacaaacc cgagcatggc atcgtgtaaa gtgttaaggt 240ccaatggcct cctgtccacg tttggcgatg taaatcctcc 280622274DNAZea mays 622cagtaaggag gttagctgtt gatgccacgc ttagagcagc tgcaccatac caaaaactgc 60gcagagagaa agaacgtgac aaaacaagaa aggttttcgt tgaaaagact gacatgagag 120ccaaaagaat ggctcgaaaa gcaggtgctc tagtcatatt tgttgtggac gctagtggta 180gcatggctct gaatcgtatg cagaatgcta aaggtgcggc gttgaagttg cttgcagaaa 240gctacaccag cagagatcag gtttcaatta ttcc 274623252DNAZea mays 623aaagcctatg cttcctaagg gtccagtaag gaggttagct gttgatgcca cgcttagagc 60agctgcacca taccaaaaac tgcgcagaga gaaagaacgt gacaaaacaa gaaaggtttt 120tgttgaaaag actgacatga gagccaaaag aatggctcga aaagcaggtg ctctagtcat 180atttgttgtg gacgctagtg gtagcatggc tctgaatcgt atgcagaatg ctaaaggtgc 240ggcgttgaag tt 252624252DNAZea mays 624aaagcctatg cttcctaagg gtccagtaag gaggttagct gttgatccca cgcttagagc 60agctccacca taccaaaaac tgcgcagaga gaaagaacgt gacaaaacaa gaaaggtttt 120tgttgaaaag actgacatga gagccaaaag aatggctcga aaagcaggtg ctctagtcat 180atttgttgtg gacgctagtg gtagcatggc tctgaatcgt atgcagaatg ctaaaggtgc 240ggcgttgaag tt 252625260DNAZea mays 625caaaaacagc gcagagagaa agaacgtgac aaaacaagaa aggtttttgt tgaaaagact 60gacatgagac ccaaaagaat ggctcgaaaa gcaggtgctc tagtcatatt tgttgtagac 120gctagtagta gcatggctct gaatcgtatg cagaatgcta aaggtgcggc gttgaagttg 180cttgcagaaa gctacaccag cagagatcag gtttcaatat tccttttcgt ggagattatc 240tgaggtttgc tccaccatca 260626260DNAZea mays 626caacccatca gaggccacgg tggccaagcg ccggagctac gcgaacacca tcagctacct 60gaccccaccg gccgagaacg ccggcctcta caaggggctc aagcagctgt cagagctcat 120ctcttcctac cagtctctca aggacaccgg gcgtggtcct cagattgtga gctccatcgt 180cagcactgca aagcagtgca acctcgacaa ggatgtcccg ctgcccgagg aaggggagga 240gtcccaccaa aggagcgtga 260627122DNAZea mays 627caaggacacc gggcgtggtc ctcagattgt gagctccatc gtcagcactg caaagcatgc 60aacctcgaca aggatgtccc cctgcctgag gaaggggagg agctcccacc aaaggagcgt 120ga 122628306DNAZea mays 628gtcgacgtgc tgctggattc cgctgcgtcg gggtggaaca cggtggagag ggacggtatc 60tccatatccc accctgctcg cttcatcctc atcggctctg gtaacccgga ggaaggggag 120ctcaggcccc agctgctgga ccggttcggg atgcacgcgc aggttggtac cgtcagggac 180gccgagctca gggtgaagat cgtggaggag agggctcgtt tcgacaggga tccgaagacg 240ttccgtgagt cgtatcatga cgagcaggag aagctccagc agcagatatc atctgcacgg 300agtaac 306629269DNAZea mays 629acctcgttga cgtgctgctg gattccgctg cgtcggggtg gaacacggtg gagagggagg 60gtatctccat atcccaccct gctcgcttca tcctcatcgg ctctggtaac ccggggaagg 120ggagctcagg ccccagctgc tggaccggtt cgggatgcac gcgcaggttg gtaccgtcag 180ggacgccgag ctcagggtga agatcgtgga ggagagggct cgtttcgaca gggatccgaa 240gacgttccgt gagtcgacca tgacgagca 269630269DNAZea mays 630caccctgctc gcttcatcct catcggctct ggtaacccgg aggaagggga gctcaggccc 60cagctgctgg accggttcgg gatgcacgcg caggttggta ccgtcaggga cgccgagctc 120agggtgaaga tcgtggagga gagggctcgt ttcgacaggg atccgaagac gttccgtgag 180tcgtaccatg acgagcagga gaagtccagc agcagatatc atctgcacgg ataacttggc 240gctgtgcaga ttgaccatga ctccgtgtc 269631433DNAZea mays 631cgtcgacctg ctcccggaca tccgcgtcgt cgtcggcgac cccttcaact ccgacccgga 60cgaccccgag gtcatgggcc ccgaggtccg ccagcgggtc ctgcaggggg acaccggcct 120ccccgtcacc accgccaaga tcaccatggt cgacctgccc ctcggcgcca ccgaggaccg 180cgtctgcggc accattgaca tcgagaaggc gctcaccgag ggcgtcaagg cgttcgagcc 240cggcctgctc gccaaggcca acaggggcat actgtacgtc gacgaggtca acctgctgga 300cgaccacctc gtcgacgtgc tgctggattc cgctgcgtcg gggtggaaca cggtggagag 360ggagggtatc tccatatccc accctgctcg cttcatcctc atcggctctg gtaacccgga 420ggaaggggag ctc 433632281DNAZea mays 632ggggcacggg gaagtccacc accgtccgct ccctcgtcga cctgctcccg gacatccgtc 60gtcgtcgtcg gcgacccctt caactccgac ccggacgacc ccgaggtcat gggccccgag 120gtccgccagc gggtcctgca gggggacacc ggcctccccg tcaccaccgc caagatcacc 180atggtcgacc tgcccctcgg cgccaccgag gaccgcgtct gcggcaccat tgacatcgag 240aaggcgctca ccgagggcgt caaggcgttc gagcccggcc t 281633273DNAZea mays 633tgcccctcgg cgccaccgag gaccgcgtct gcggcaccat tgacatcgag aaggcgctca 60ccgagggcgt caaggcgttc gagcccggcc tgctcgccaa ggccaacagg ggcatactgt 120acgtcgacga ggtcaacctg ctggacgacc acctcgtcga cgtgctgctg gattccgctg 180cgtcggggtg gaacacggtg gagagggagg gtatctccat atcccaccct gctcgcttca 240tcctcatcgg ctctggtaac ccggaggaag ggg 273634227DNAZea mays 634agatcggcgg cgtcatgatc atgggcgaca ggggcacggg gaagtccacc accgtccgct 60ccctcgtcga cctgctcccg gacatccgcg tcgtcgtcgg cgaccccttc aactccgacc 120cggacgaccc cgaggtcatg ggccccgagg tccgccagcg ggtcctgcag ggggacaccg 180gcctccccgt caccaccgcc aagatcacca tggtcgacct gcccctc 227635372DNAZea mays 635cccacgcgtc cgggcaagtc gtcaatgttg ccaacaacct cagcaagata cttggtttcg 60gcctgtcgga accatgggtg cagtacctgt ccacgaccaa gttcgtcaga gcggacagag 120agaagatgag ggttctgttt gggttcttgg gggagtgcct gaggctcgtc gtgcaagaca 180acgagctggg aagcttgaag cttgccctcg agggaagcta cgtcgagcct ggccctggcg 240gcgacccgat ccgtaacccg aaggtgctcc cgacagggaa gaacatccac gctctcgatc 300cgcaggccat cccaaccacg gctgccttga agagcgccaa gatcgtcgtg taccgtctcc 360tggagaggca ga 372636263DNAZea mays 636gttcgtcaga gcggacagag agaagatgag ggttctgttt gggttcttgg gggagtgcct 60gacggtcgtc gtgcaagaca acgagctggg aagcttgaag cttgccctcg agggaagcta 120cgtcgagcct ggccctggcg gcgacccgat ccgtaacccg aaggtgctcc cgacagggaa 180gaacatccac gctctcgatc cgcaggccat cccaaccacg gctgccttga agagcgccaa 240gatcgtcgtg gaccgtctcc tgg 263637272DNAZea mays 637cccacgcgtc cggttgccaa caacctcagc aagatacttg gtttcggcct gtcggaacca 60tgggtgcagt acctgtccac gaccaagttc gtcagagcgg acagagagaa gatgagggtt 120ctgtttgggt tcttggggga gtgcctgatg ctcgtcgtgc aagacaacga gctgggaagc 180ttgaagcttg ccctcgaggg aagctacgtc gagcctggcc ctggcggcga cccgatccgt 240aacccgaagg tgctcccgac agggaagaac at 272638273DNAZea maysunsure at all n locations 638gtttgggttc ttgggggagt gcctgangnt cgtcgtgcan gacaangagc ttggaatctt 60gaatcttgcc ctcgagggaa gctacgtcga gcctggccct ggcggcgacc cgattncgta 120acccgaaggt gctcccgaca ggaagaacat ctangctctt nnatccgcan gccatcccaa 180ccacggctgc cttgaagagc gncaagatcg tcgtggaccg tctcctggag aggcagaagg 240ctgacaatgg nggcaagtac cctgagacgg tcg 273639301DNAZea mays 639acttgctgaa gcacatagag gtgttcttta tgttgatgaa ataaatctat tggatgatgg 60cataagcaat ctacttctga atgtcttgac ggagggagtt aacattgtgg aaagagaggg 120cattagcttt cgccatccct gcaaaccact tctaattgct acttacaatc cagaggaagg 180gtctgtacgt gaacacttgc ttgatcgtat tgcaattaat ttaagtgctg atcttccaat 240gagttttgat gaccgcgttg aagcagtgga tattgcaaca cggtttcagg agtctagcaa 300a 301640307DNAZea mays 640ggtgttcttt atgttgatga aataaatcta ttggatgatg gcataagcaa tctacttctg 60aatgtcttga cggagggagt taacattgtg gaaagagagg gcattagctt tcgccatccc 120tgcaaaccac ttctaattgc tacttacaat ccagaggaag gatctgtacg tgaacacttg 180cttgatcgta ttgcagttaa tttaagtgct gatcttccaa tgagttttga tgaccgcgtt 240gaagcagtgg atattgcaac acggtttcag gagtctaggc aagaagtttt caaattggtg 300gaagaaa 307641278DNAZea maysunsure at all n locations 641tgttgatgaa ataaatctat tggatgatgg cataagcaat ctacttctgn atgtcgtgac 60ggagggagtt aacattgtgg aaagagaggg gattagcttt cgccatccct gcaaaccact 120tctaattgct acttacaatc

cagaggaagg atctgtacgt gaacactctg ctgatcgtat 180tgcattaatt aagtgctgat cagcaatgag tttgatgacg cgttgaacat ggatatcaca 240ccggttcaga gctacaagaa tttcaatcgt ggagaaaa 278642426DNAZea mays 642cccacgcgtt cgcccacgcg ttcgcggtga caagggtgtt ctcgaacgca tcaggctggt 60actcgtccaa cgtgaacctg gccgtggaga acgcgtcatg gaccgacgag aagcagctcc 120aggacatgta cctgagccgc aagtccttcg cgttcgacag cgacgcccca ggggcaggca 180tgaaggagaa gcgcaaggcg ttcgagctcg ccctggcgac ggcggacgcc acgttccaga 240acctcgactc gtcggagatc tcgctgacgg acgtgagcca ctacttcgac tcggacccga 300ccaagctcgt gcaggggctg cgcaaggacg ggcgggcgcc gtcctcgtac atagccgaca 360ccaccacggc gaacgcccag gtgaggacgc tgtcggagac ggtgcgcctc gacgcgagga 420ccaagc 426643312DNAZea mays 643ccgcgtgtcg ctaagggagg cggcgacaag ggtgttctcg aacgcatcac gctcctactc 60gtccaacgtg aacctggccg tggagaacgc gtcatggacc gacgagaagc agctccagga 120catgtacctg acccgcaagt ccttcgcgtt cgacagcgac gccccagggg caggcatgaa 180ggagaagcgc aaggcgttcg acctcgccct ggcgacggcg gacgccacgt tccagaacct 240cgactcgtcg gagatctcgc tgacggacgt gagccactac ttcgactcgg acccgaccaa 300gctcgtgcag gg 312644287DNAZea mays 644acgtgagcca ctacttcgac tcggacccga ccaagctcgt gcaggggctg cgcaaggacg 60ggcgggcgcc gtcctcgtac atagccgaca ccaccacggc gaacgccagg tgaggacgct 120gtcggagacg gtgcgcctcg acgcgaggac caagctgctg aaccccaagt ggtacgaggg 180gatgatgaag agcgggtacg agggggtcag ggagatcgag aagcggctca ccaacaccgt 240cgggtggagc gccacgtctg ggcaggtcga caactgggtc tacgagg 287645279DNAZea mays 645gtacctgagc cgcaagtcct tcgcgttcga cagcgacgcc ccaggggcag gcatgaagga 60gaagcgcaag gcgttcgagc tcgccctggc gacggcggac gccacgttcc agaacctcga 120ctcgtcggag atctcgctga cggacgtgag ccactacttc gactcggacc cgaccaagct 180cgtgcagggg ctgcgcaagg acgggcgggc gccgtcctcg tacatagccg acaccaccac 240ggcgaacgcc aggtgaggac gctgtcggag acggtgcgc 279646280DNAZea mays 646aagatggtgg ccgaactgga cgagccagca gagatgaact acgtgcgaat accccaggag 60taggcggagg agctcggcgt gtcgctaagg gaagcggcga caagggtgtt ctcgaacgca 120tcaggctcct actcgtccaa cgtgaacctg gcggtggaga acgcgtcatg gaccgacgat 180aagcagctcc aggacatgta cctgagccgc aagtccttcg cgttcgacag cgacgcccct 240ggggcaggca tgaaggagaa gcgcaaggcg ttcgagctcg 280647213DNAZea mays 647ggcgacggcg gacgccacgt tccagaacct cgactcgtcg gagatctcga tgacggacgt 60gagccactac ttcgactcgg acccgaccaa gctcgtgcag gggctgcgca aggacgggcg 120ggcgccgtcc tcgtacatag ccgacaccac cacggcgaac gcccaggtga ggacgctgtc 180ggagacggtg cgcctcgacg cgaggaccaa gct 213648166DNAZea mays 648aagcacgccc aggagcaggc ggaggagctc ggcgtgtcgc taagggaggc ggcgacaagg 60gtgttctcga acgcatcagg ctcctactcg tccaacgtga acctgacggt ggagaacgcg 120tcatggaccg acgagaagca gctccaggac atgtacctga gccgca 166649449DNAZea mays 649gggatgatga agagcgggta cgagggggtc agggagatcg agaagcggct caccaacacg 60cgtcgggtgg agcgccacgt ctgggcaggt cgacaactgg gtctacgagg aggccaactc 120cacgttcatc gaggacgagg cgatgaggaa gaggctcatg gacaccaacc ccaattcgtt 180caggaagttg gtgcagacct tcctggaagc cagtggcaga ggctactggg agacaacgga 240ggagaacctg gacaggctca gggagctcta ttcggaggtt gaagacaaga ttgaggggat 300tgacaggtaa attgatttgc cagatcggtc ggccgatcgg ttccagcatt caacccataa 360cgagcttgga actcttctgc ctcattggga ctcttgtaca atgtctgggt gtgtgattta 420tatatatata aaagtgtaac atgtaatac 449650305DNAZea mays 650cgagaagcgg ctcaccaaca ccgtcgggtg gagcgccacg tctgggcagg tcgacaactg 60ggtctacgag gaggccaact ccacgttcat cgaggacgag gcgatgagga agaggctcat 120ggacaccaac cccaattcgt tcaggaagtt ggtgcagacc ttcctggaag ccagtggcag 180aggctactgg gagacaacgg aggagaacct ggacaggctc agggagctct attcggaggt 240tgaagacaag attgagggga ttgacaggta aattgatttg ccagatcggt cggccgatcg 300gttcc 305651270DNAZea mays 651gacgcgagga ccaagctgct gaaccccaag tggtacgagg ggatgatgaa gagcgggtac 60gagggggtca gggagatcga gaagcggctc accaacaccg tcgggtggag cgccacgtct 120gggcaggtcg acaactgggt ctacgaggag gccaactcca cgttcatcga ggacgaggcg 180atgaggaaga ggctcatgga caccaacccc aattcgttca ggaagttggt gcagaccttc 240ctggaagcca gtggcagagg ctactgggag 270652440DNAZea maysunsure at all n locations 652cattgttcag ctgccggctc agtatctgag actcgtgggt cgtcacaagc ctctacactg 60acgtcctact aggacgaggc gatgaggaag aggctcatgg acaccaaccc caattcgttc 120aggaagttgg tgcagacctt cctggaagcc agtggcagag gctactggga gacaacggag 180gagaacctgg acaggctcag ggagctctat tcggaggttg aagacaagat tgaggggatt 240gacaggtaaa ttgatttgcc agatcggtcg gccgatcggt tccagcattc aacccataac 300gagcttggaa ctcttctgcc tcattgggac tcttgtacaa tgtctgggtg tgtgatttat 360atatatataa aaagttgtaa catgtaatac tggaggatac aatatttaac anagagggtg 420gcggttgttc catccaaaac 440653213DNAZea mays 653tgcagatccg gacattatcc gtcttcctag gctctttcgc tttctgcaga agccacttgc 60aaaattcata tcagaagtga gagcaccaaa aagtaaggaa ggttatgcat ccataggtgg 120cggttctcct ctacgacaaa ttactgatgc acaggctgaa gcactgaggg aggcattaca 180tgggaaagat gccctgccaa cgtgtatgtt gga 213654261DNAZea mays 654cccacgcgtc cgggtaccct ttcacagaag aggccattga tcaaattaaa aaggataaga 60ttaccaagct cgttgttctt cccctttacc ctcagtactc catatcaaca agtgggtcaa 120gcattcgtgt tctccaagac attgtcaagg aagattcata tttttctggt ttgccaattt 180ccattattga atcatggtac caacgagatg gctatgtgaa atcaatgtct gacctaattg 240aaaaggagct ctcggccttc t 261655291DNAZea mays 655tgagatccag aggaatctta aatggtcaca ctttggcgta tcagagtcgg gtgggaccag 60ttcaatggct gaagccatat actgatgaag ttttagtaga aattggtcag aacggtgtga 120agagcctcct ggctgttcca gtaagcttcg tgagcgagca cattgagaca ctggaagaaa 180tagacatgga gtacaaggag ttggctctgg aatcaggcat tgagaactgg ggccgggtcc 240ctgctcttgg atgcacttcg acgttcatct ccgacttgca gatgcggttg t 291656275DNAZea mays 656actgctagca gcatacgact cgaagcgcga tgagctccct ccaccggtaa tcgtgtggga 60gtggggctgg acaaagagcg cggagacctg gaatagccgt gcggcgatgc tggccgtgct 120ggctctcctg gtgctggaag tgaccaacgg cgaagggttc ctgcatcaat ggggaatcct 180gcctctgttc cgctgagccg acaattctgt tcatgatggg gtcataattt tgctgcagcc 240gaaggaagtt ttgaacttct gatgctgtat atgaa 275657261DNAZea maysunsure at all n locations 657atcaagagga atcttagata gtcatacttt ggcgtaccag aatcgggtgg agctagttca 60atggctgaag ctatatactg atgaagtatt agtagaactt ggtgaaaagg gtgtgaagag 120cctactggct gttacagtaa gccttgagag taaagacatc gagacattgg aagaaattga 180catggagtac aaggagttgg ctctggaatc aggcatcaag aactggggtc gggttcctgc 240tctgatnnac acttcaacat t 261658398DNAZea mays 658acggacgcgt gggtttagca taacacgggg tgcatgcaca tgtatccgat tccctgcatc 60actcacacct cactttttct gctaaattgt ggcagtggtg ataattgata tgcatagact 120gtacttattt aatgactatg aaataccatt taacatagct attgtgcctg acagggtaaa 180tctaccaagg acacacatag ttaagccttg ctcagctgac gactgctaag gaatttctgt 240taagtgcagt ttggggggtc ttctcaacca ttgcttgact taaggcaaca cattagagga 300tattcatcag catcagaggc aattcttccc aatctgattt gagaaaaaaa tttgttggca 360acgaaaaatt agtgttttct tgctgaatct tggggggc 398659356DNAZea mays 659gctttgatca tgggggagtt aagatcaaga ggaatcttaa atagtcacac tttggcgtac 60caggtaaatg ctattaaaat ttggtaggta attgtttcac taacaacgga gttgtgccct 120tatgttttaa tgatcacctt gtaagaacac taggaatgga aactgccaag ttatataggc 180ttcaggagtt accagttcct taattttcca ggtcaccatt aactagtgtt aacatttatt 240gtacacgcag agtcgggtgg ggccagttca atggctgaag ccatatactg atgaagtttt 300agtagaactt ggtcaaaagg gtgttaagag cctcctggct gttccagtaa gctttg 356660266DNAZea mays 660cccacgcgtc cgaaagatgt tcctgccaac gtgtatgttg gaatgcggta ttggcatccc 60ttcactgaag aagccataga acaaataaaa cgggatggaa tcacgaaact tgttgtgttg 120cctctatacc ctcagttctc catatcaact agtggttcaa gtctccgttt attggagagc 180atattcagag aggatgagta tctcgtgaat atgcaacata cagttatacc ttcctggtac 240caacgtgaag gatatatcaa ggctat 266661260DNAZea mays 661cggacgcgtg gcgcgacgcg tgggcggacg cgtgggcgga cggtggggaa agatgttcct 60gccaacgtgt atgttggaat gcggtattgg catccctatc actgaagaag ccatagaaca 120aacaaaacgg gatgcaatca cgaaacttgt tgtgttgcct ctataccctc agttctccat 180atcaactagt ggttcaagtc tccgtttatt ggagagcata ttcagagagg atgagtatct 240cgtgaatatg caacatacag 260662195DNAZea mays 662cccacgcgtc cgcccacgcg tccgcccacg cgtccgccca cgcgtccgat ggaatcacga 60aacttgttgt gttgcctcta taccctcagt tctccatatc aactagtggt tcaagtctcc 120gtttattgga gagcatattc agagaggatg agtatctcgt gaatatgcaa catacagtta 180taccttcctg gtacc 195663430DNAZea maysunsure at all n locations 663gccgccgttg ggccttttgc cggcgacggg aacccatcac accaggtcat ggggcaaaac 60aacctccaca agttttactg gttctaccac caaacatgag cagagcttgc atggaaatgt 120taagccgttg caattggcgg caaatgaatc ctctcgtttg gcttacagaa gtccagcact 180taaaaaccag tggaatcttc ctgctagttc ttcctccact aatgtggtta ccacctttga 240tgataacgaa cacgtgtctt ccagtgttat tgaagaaaaa gttggagtac tgttattaaa 300ccttggtggt ccagagacac ttgacgatgt tcaaccattt ttattcaacc tatttgctga 360tccagatatc attcgactcc ctangctctt caagtttcct cnaagacact gggcaaacnt 420ntatttaatt 430664199DNAZea mays 664aaacaacctc cacaagtttt actggttcta ccaccaaaca tgagcagagc ttgcatggaa 60atgttaagcc gttgcaattg gcggcaaatg aatcctctcg tttggcttac agaagtccag 120cacttaaaaa ccagtggaat cttcctgcta gttcttcctc cactaatgtg gttaccacct 180ttgatgataa cgaacacgt 199665443DNAZea mays 665gccacgtttg gtagttgcta cttgctacac cggaggaaga agaacaagta gtgcttttct 60tctcttgtca cgttcacggg gcggccgatc gaccgttcac ctcgcccgac ggcccaagca 120gcccatgtct tcgtcgggcc cctccccggc gacgggaatc cacgcgtcgc cgccgttggg 180ccttttgccg gcgacgggaa cccatcacac caggtcatgg ggcaaaacaa cctccacaag 240ttttactggt tctaccacca aacatgagca gagcttgcat ggaaatgtta agccgttgca 300attggcggca aatgaatcct ctcgtttggc ttacagaagt ccagcactta aaaaccagtg 360gaatcttcct gctagttctt cctccactaa tgtggttacc acctttgatg ataacgaaca 420cgtgtcctcc agtgttattg aag 443666304DNAZea mays 666gagactccat atcaacaagt agcatatttt ttactaagaa gaagagaagg gaagattcat 60atttttctgg cttgccaatc tccattatcg aatcatggta ccaacgtgat ggctatgtga 120aatcaatggc tgacctaatt gaaaaagagc tatctgcctt ttccaatcct gaagaggtaa 180tgatatgctt cagtgcacat ggtgtgccac ttacctatgt tcaggatgct ggagatcctt 240acagagatca gatggaggat tgtatttctg tgatcatggg ggagctgaga tccagaggaa 300tctt 304667256DNAZea mays 667ttcgtgttct ccgaaatgtt gtcaagggag attcatattt ttctggcttg gcaatctcca 60gtatcgaatc atggtagcaa cgtgatggct atgtgaaatc agtggctgac ctgattgaga 120aagaggtatc tgccttttcc agtcctgaag aggtagtgat attcttcagt gcacatagtg 180tgccacttag ctatgtgcag gatgctggag atccttacag agatcagatg gatgattgta 240tttctttgat cgtggg 256668263DNAZea mays 668agaggttatg atattcttca gtgcacatgg tgtgccactt acctatgttg aggatgctgg 60agatccttac agagatcaga tggaggattg tattgctttg atcatggggg agttaagatc 120aagaggaatc ttaaatagtc acactttggc gtaccagagt cgggtggggc cagttcaatg 180gctgaagcca tatactgatg aagttttagt agaacttggt caaaagggtg tgaagagcct 240catggctgtt ccagtaagct ttg 263669266DNAZea mays 669agaggttatg atattcttca gtgcacatgg tgtgccactt acctatgttg aggatgctgg 60agatccttac agagatcaga tggaggattg tattgctttg atcatggggg agttaagatc 120aagaggaatc ttaaatagtc acactttggc gtaccagagt cgggtggggc cagttcaatg 180gctgaagcca tatactgatg aagttttagt agaacttggt caaaagggtg tgaagagcct 240cctggctgtt ccagtaagct ttgtga 266670276DNAZea mays 670atctgccttt tccaatcctg aagaggtaat gatattcttc agtgcacatg gtgtgccact 60tacctatgtt caggatgctg gagatcctta cagagatcag atggaggatt gtatttcttt 120gctcatgggg gagctgagat ccagaggaat cttaaatggt cacactttgg cgtatcagag 180tcgggtggga ccagttcaat ggctgaagcc atatactgat gaagttttag tagaacttgg 240tcagaacggt gtgaagagcc tcctggctgt tccagt 276671307DNAZea mays 671ctgttattaa accttggtgg tccagagaca cttgacgatg ttcaaccatt tttattcaac 60ctatttgctg atccagatat cattcgactc cctaggctct tcaggtttct tcaaagacca 120ctggccaaac ttatttctac ttttagagct cctaagagta aagaagggta tgcttcaatg 180gtggtgggtc gccgttaagg aaaattactg atgaacaggc gaatgctttg aagattgccc 240tggaaaagaa aaaattgaac gcaaacatat atgttgggat gcggtattgg taccctttca 300cagaaga 307672310DNAZea mays 672ctgttattaa accttggtgg tccagagaca cttgacgatg ttcaaccatt tttattcaac 60ctatttgctg atccagatat cattcgactc cctaggctct tcaggtttct tcaaagacca 120ctggccaaac ttatttctac ttttagagct cctaagagta aagaagggta tgcttcaatt 180ggtggtgggt cgccgttaag gaaaattact gatgaacagg cgaatgcttt gaagattgcc 240ctggaaaaga aaaaattgaa cgcaaacata tatgttggga tgcggtattg gtaccctttc 300acagaagagg 310673122DNAZea mays 673cccacgcgtc cggcttcaat cggtggtggg tcaccattga ggaaaattac tgatgagcag 60gcaaatgctt tgaagattgc tctggaaaag aaaaaattga acgcaaatat atatgttggg 120at 122674431DNAZea maysunsure at all n locations 674cggacgcgtg ggttggacca gtggaatggc tgaaaccgta cactgatgag acagtgatgg 60agcttgggca gaaaggggta aagagcctgc ttgctgttcc cattagtttt gttagcgaac 120acattgaaac tttggaagaa atcgatgtgg agtacaaaga gttggctttg gaatctggca 180tcaagcactg gggacgggtt ccagcactag gttgcgaacc cacattcatt tcggatcttg 240ctgatgctgt tattgaaagc ctaccttatg ttggcgcaat ggcagtttcc aatcttgagg 300ctcggcagtc tctcgtaccc ctcgggagcg tggaggagct gctagcagca tacgactcga 360agcgcgatga gctccctcca ccggtaatcg tgtgggagtg gngctggaca aagagcgcgg 420agacctggaa t 431675298DNAZea mays 675agactggaaa aaagaggaat aacaaatccg tgcatacttg cttatcagag ccgagttgga 60ccagtggaat ggctgaaacc gtacactgat gagacaatta ttgagcttgg gcagaaaggg 120gtaaagagcc tgcttgctgt tcccattagt tttgttagcg aacacattga aactttggaa 180gaaatcgatg tggagtacaa agagttggct ttggaatctg gcatcaagca ctggggacgg 240gttccagcac taggttgcga acccacattc atttcggatc ttgctgatgc tgttattg 298676308DNAZea mays 676gagacgcgtg gcggacgcgt gggcggacgc gtggggccga gttggaccag tggaatggct 60gaaaccgacc actgatgaga ctattattga gattgggcag aaaggggtaa agagcctgct 120tgctgttccc attagttttg ttagcgaaca cattgaaact ttggaagaaa tcgatgtgga 180gtacaaagag ttggctttgg aatctggcat caagcactgg ggacgggttc cagcactagg 240ttgcgaaccc acattcattt cgtatcttgc tgatgctgtt attgaaacct accttatgtt 300ggcgcatg 308677174DNAZea mays 677cccacgcgtc cggcttgggc agaaaggggt aaagagcctg cttgctgttc ccattagttt 60tgttagcgaa cacattgaaa ctttggaaga aatcgatgtg gagtacaaag agttggcttt 120ggaatctggc atcaagcact ggggacgggt tccagcacta ggttgcgaac ccac 174

Claims

1-9. (canceled)

10. A transformed plant comprising a nucleic acid molecule which comprises:(a) an exogenous promoter region which functions in a plant cell to cause the production of an mRNA molecule; which is linked to;(b) a structural nucleic acid molecule, wherein said structural nucleic acid molecule comprises a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof, or which is linked to(c) a 3' non-translated sequence that functions in said plant cell to cause the termination of transcription and the addition of polyadenylated ribonucleotides to said 3' end of said mRNA molecule.

11. The transformed plant according to claim 10, wherein said nucleic acid sequence is the complement of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677.

12. The transformed plant according to claim 10, wherein said nucleic acid sequence is in the antisense orientation of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677.

13. The transformed plant according to claim 10, wherein said nucleic acid sequence shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

14. The transformed plant according to claim 13, wherein said nucleic acid sequence shares between 100% and 98% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

15. The transformed plant according to claim 14, wherein said nucleic acid sequence shares between 100% and 99% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

16. The transformed plant according to claim 15, wherein said nucleic acid sequence shares 100% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

17. A transformed seed comprising a transformed plant cell comprising a nucleic acid molecule which comprises:(a) an exogenous promoter region which functions in said plant cell to cause the production of an mRNA molecule; which is linked to;(b) a structural nucleic acid molecule, wherein said structural nucleic acid molecule comprises a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof, which is linked to(c) a 3' non-translated sequence that functions in said plant cell to cause the termination of transcription and the addition of polyadenylated ribonucleotides to said 3' end of said mRNA molecule.

18. The transformed seed according to claim 17, wherein said nucleic acid sequence is the complement of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677.

19. The transformed seed according to claim 17, wherein said exogenous promoter region functions in a seed cell.

20. The transformed seed according to claim 17, wherein said nucleic acid sequence shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

21. The transformed seed according to claim 20, wherein said nucleic acid sequence shares between 100% and 98% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

22. The transformed seed according to claim 21, wherein said nucleic acid sequence shares between 100% and 99% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

23. The transformed seed according to claim 22, wherein said nucleic acid sequence shares 100% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

24. A method of growing a transgenic plant comprising(a) planting a transformed seed comprising a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof, and(b) growing a plant from said seed.

25. A substantially purified nucleic acid molecule comprising a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 677 and complements thereof.

26. The substantially purified nucleic acid molecule of claim 25, wherein said nucleic acid molecule encodes a maize protein or fragment thereof.

27. The substantially purified nucleic acid molecule of claim 25, wherein said nucleic acid molecule encodes a soybean protein or fragment thereof.

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