Patent application title: METHOD FOR SPEEDING UP PLANT GROWTH AND IMPROVING YIELD BY ALTERING EXPRESSION LEVELS OF KINASES AND PHOSPHATASES
Inventors:
Boon Leong Lim (Hong Kong, HK)
IPC8 Class: AC12N1582FI
USPC Class:
800290
Class name: Multicellular living organisms and unmodified parts thereof and related processes method of introducing a polynucleotide molecule into or rearrangement of genetic material within a plant or plant part the polynucleotide alters plant part growth (e.g., stem or tuber length, etc.)
Publication date: 2016-03-24
Patent application number: 20160083744
Abstract:
Transgenic plants having increased growth rate and increase yield are
disclosed, and methods for making the same. In one embodiment, the method
comprises: transforming a plant or plant cell with a nucleic acid
molecule comprising a plant kinase and/or phosphatase gene selected from
NG6, NG21, NG24, NG28, and NG32, and over-expressing said kinase and/or
phosphatase gene in the plant or plant cell.Claims:
1. A method to make a transgenic plant having increased rate of plant
growth and/or elevated plant yields comprising: a) introducing a gene
expression construct into a plant or plant cell, wherein the construct
comprises a nucleic acid molecule encoding a phosphatase or kinase and
the nucleic acid molecule comprises: i) a sequence having at least 85%
identity with SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25,
27, 29, 31, 33, 35, 37, 39, 41, 45, 47, 49, 93, 95, 97, 99, or 101; or
ii) a sequence encoding a polypeptide having at least 85% identity with
SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34,
36, 38, 40, 42, 46, 48, 50, 94, 96, 98, 100, or 102; wherein the
sequence is operatively linked to upstream and downstream regulatory
components comprising a heterologous promoter; and b) overexpressing the
nucleic acid molecule in the plant or plant cell, wherein the promoter
drives the overexpression of the nucleic acid molecule.
2. The method of claim 1, wherein the nucleic acid molecule comprises: i) a sequence selected from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 45, 47, 49, 93, 95, 97, 99, or 101; or ii) a sequence encoding a polypeptide selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 46, 48, 50, 94, 96, 98, 100, or 102.
3. The method of claim 1, further comprising regenerating, from said transformed plant or plant cell, a plant having enhanced growth and/or yield.
4. The method of claim 1, wherein plant growth rate is increased.
5. The method of claim 1, wherein plant yield is increased.
6. The method of claim 1, wherein the plant is of a genus selected from the group consisting of: Asparagus, Bromus, Hemerocallis, Hordeum, Lolium, Panicum, Pennisetum, Saccharum, Sorghum, Trigonella, Triticum, Zea, Antirrhinum, Arabidopsis, Arachis, Atropa, Brassica, Browallia, Capsicum, Carthamus, Cichorium, Citrus, Chrysanthemum, Cucumis, Datura, Daucus, Digitalis, Fragaria, Geranium, Glycine, Helianthus, Hyscyamus, Ipomoea, Latuca, Linum, Lotus, Majorana, Malva, Gossypium, Manihot, Medicago, Nemesia, Nicotiana, Onobrychis, Pelargonium, Petunia, Ranunculus, Raphanus, Salpiglossis, Senecio, Sinapis, Solanum, Trifolium, Vigna, and Vitis.
7. The method of claim 1, wherein the plant is of a species of the genus Brassica.
8. The method of claim 1, wherein the plant is of a species selected from the group consisting of: Glycine max, Zea mays, Solanum tuberosum, Panicum virgatum, Medicago sativa, and Brassica napus.
9. The method of claim 1, wherein the plant cell is a seed, stem, shoot, or root cell.
10. (canceled)
11. The method of claim 1, wherein plant weight, total weight of leaf or seed, total number of inflorescence, carbon metabolism, level of carbohydrate, amino acid, lipid production or seed yield per plant are increased; or bolting time is earlier, when compared to a wild-type plant of the same species cultivated under the same conditions.
12. A transgenic plant cell, comprising: a gene expression construct, wherein the construct comprises: i) a nucleic acid molecule comprising a sequence having at least 85% identity with SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 45, 47, 49, 93, 95, 97, 99, or 101, wherein said nucleic acid molecule is overexpressed in the transgenic plant cell when compared to plant cells of the same type in a wild-type plant of the same species; or ii) a nucleic acid molecule that encodes a protein comprising a sequence having at least 85% identity with SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 11, 16, 48, 50, 94, 96, 98, 100, or 102, wherein said nucleic acid molecule is overexpressed in the transgenic plant cell when compared to plant cells of the same type in a wild-type plant of the same species; wherein the sequence is operatively linked to upstream and downstream regulatory components comprising a heterologous promoter that drives the overexpression of the nucleic acid molecule in the plant cell.
13. The transgenic plant cell of claim 12, comprising: i) a nucleic acid molecule comprising the sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 45, 47, 49, 93, 95, 97, 99, or 101, wherein said nucleic acid molecule is overexpressed in the transgenic plant cell when compared to plant cells of the same type in a wild-type plant of the same species; or ii) a nucleic acid molecule that encodes a protein comprising the sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100, or 102.
14. The transgenic plant cell of claim 12, wherein the plant cell comprises a nucleotide sequence having at least 85% identity with SEQ ID NO: 1, 3, 5, 7, 9.
15. The transgenic plant cell of claim 12, wherein the plant cell comprises a nucleotide sequence encoding a sequence having at least 85% identity with SEQ ID NO:2, 4, 6, 8, or 10.
16. The transgenic plant cell of claim 12, wherein said plant cell is of a monocotyledonous species.
17. The transgenic plant cell of claim 12, wherein said plant cell is of a dicotyledonous species.
18. The transgenic plant cell of claim 12, wherein the plant cell is of a genus selected from the group consisting of: Asparagus, Bromus, Hemerocallis, Hordeum, Lolium, Panicum, Pennisetum, Saccharum, Sorghum, Trigonella, Triticum, Zea, Antirrhinum, Arabidopsis, Arachis, Atropa, Brassica, Browallia, Capsicum, Carthamus, Cichorium, Citrus, Chrysanthemum, Cucumis, Datura, Daucus, Digitalis, Fragaria, Geranium, Glycine, Helianthus, Hyscyamus, Ipomoea, Latuca, Linum, Lotus, Majorana, Malva, Gossypium, Manihot, Medicago, Nemesia, Nicotiana, Onobrychis, Pelargonium, Petunia, Ranunculus, Raphanus, Salpiglossis, Senecio, Sinapis, Solanum, Trifolium, Vigna, and Vitis.
19. The transgenic plant cell of claim 12, wherein the plant cell is of a species of the genus Brassica.
20. A transgenic plant, comprising the transgenic plant cell of claim 12.
21. The transgenic plant of claim 20, wherein plant weight, total weight of leaf or seed, total number of inflorescence, carbon metabolism, level of carbohydrate, amino acid, lipid production, or seed yield per plant are increased; or bolting time is earlier, when compared to a wild-type plant of the same species cultivated under the same conditions.
22. The method of claim 11, wherein the seed yield per plant is increased by approximately 30% -50% as compared with a wild-type plant of the same species.
23. The method of claim 11, wherein the bolting time per plant is earlier by approximately 19% as compared with a wild-type plant of the same species.
24. The transgenic plant of claim 21, wherein the seed yield is increased by approximately 30%-50% as compared with a wild-type plant of the same species.
25. The transgenic plant of claim 21, wherein the boiling time is earlier by approximately 19% as compared with a wild-type plant of the same species.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 61/482,467, filed May 4, 2011, which is hereby incorporated by reference in its entirety.
1. INTRODUCTION
[0002] Described herein are methods for speeding up plant growth and/or elevating plant yields by altering the expression levels of plant kinases and phosphatases. Also described therein are the use of plant kinases and phosphatases, and their respective protein products, as well as fragments, derivatives, homologues, and variants thereof.
2. BACKGROUND OF THE INVENTION
[0003] Purple acid phosphatases (PAPs) catalyze the hydrolysis of a wide range of activated phosphoric acid mono- and di-esters and anhydrides (Klabunde et al., 1996). The PAP proteins are characterized by seven conserved amino acid residues (shown in bold face) in the five conserved motifs XDXX, XDXXY, GNH(D/E), XXXH, XHXH, which are involved in the coordination of the dimetal nuclear center (Fe3+-Me2+) in the active site (Li et al., 2002), where Me is a transition metal; Me2+ is mostly found to be Fe2+ in mammals, and Zn2+, or Mn2+ in plants (Klabunde and Krebs, 1997; Schenk et al., 1999).
[0004] Multiple PAP-like sequences are present in plant genomes. In the Arabidopsis genome, twenty-nine potential PAP genes have been identified based on sequence comparison. Most of the functions of characterized plant PAPs are related to phosphorus metabolism. None of the plant PAPs that had been functionally or biochemically characterized carry any transmembrane motif. In addition, no AtPAPs or any other plant PAPs had been discovered to affect sugar signalling and carbon metabolism in plants. Overexpression of AtPAP2 in Arabidopsis, a PAP with a C-terminal motif, can significantly speed up plant growth, increase sugar content in plants and improve seed yield (U.S. Patent Application Publication No. 2010/0159065).
3. SUMMARY
[0005] In one aspect, provided herein are methods that speed up or increase the rate of plant growth and elevate plant yields by altering the expression levels of plant kinases and phosphatases. Kinases and phosphatases, and their respectively encoded protein products, as well as fragments, derivatives, homologues, and variants thereof, are disclosed. Methods for introducing these genes into plants to speed up or increase the growth rate of plants, and to increase yield of plants, are provided. The kinases and phosphatases of the present invention are selected from the results of a microarray study. Surprisingly, it is discovered that phosphatases (such as NG6) and kinases (such as NG21, NG24, NG28, and NG32) have growth-promoting effects.
[0006] Provided herein, a microarray study was carried out to compare the gene expression profiles of the AtPAP2 overexpression lines, AtPAP2 T-DNA (mutant) line, and the wild-type plants. The results showed that expression levels of a number of genes are significantly altered (upregulated or downregulated) in AtPAP2 overexpression lines, when compared to the wild-type. Among these genes, a number of phosphatases and kinases were selected and analyzed using transgenic studies in Arabidopsis.
[0007] At least in part, the present inventors discover that altering the expression levels of plant phosphatases (such as NG6) and kinases (such as NG21, NG24, NG28 and NG32) in plants resulted in rapid plant growth and higher yield. In one aspect, provided herein are methods of producing plants with enhanced growth and/or yield. In one embodiment, the method comprises: transforming a plant or plant cell with a nucleic acid molecule comprising a plant kinase and/or phosphatase gene selected from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101, and over-expressing said kinase and/or phosphatase gene in the plant or plant cell. In one embodiment, provided herein are methods of regenerating, from said transformed plant or plant cell, a plant having enhanced growth and/or yield.
[0008] In one embodiment, the method comprises: transforming a plant or plant cell with a nucleic acid molecule comprising a plant kinase and/or phosphatase having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101, and over-expressing said kinase and/or phosphatase gene in the plant or plant cell.
[0009] In certain embodiments, the method comprises transforming a plant or plant cell with a nucleic acid molecule comprising a plant kinase and/or phosphatase having a nucleic acid fragment from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101. In certain embodiments, the nucleic acid fragment encode a peptide that has the same activity as a peptide encoded by SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101.
[0010] In certain embodiments, the activity is a kinase and/or phosphatase activity. In certain, embodiments, the method comprises transforming a plant or plant cell with a nucleic and molecule comprising a plant kinase and/or phosphatase having a variant from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101.
[0011] In certain embodiments, the variant has 1-5, 6-10, 11-20, 21-30, 31-40, 41-50, 50-70, 71-80, 81-100 nucleic acid deletion, substitution or insertion in the sequence as compared to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101. In certain embodiments, the variants encode a peptide that has the same activity as a peptide encoded by SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101. In certain embodiments, the activity is a kinase and/or phosphatase activity.
[0012] Provided herein are transgenic plants with enhanced growth and/or yield. In certain embodiments, the transgenic plant comprises a nucleic acid molecule selected from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101, wherein said nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions.
[0013] In certain embodiments, the transgenic plant comprises a nucleic acid molecule having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101, wherein said nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions.
[0014] In certain embodiments, the transgenic plant comprises a nucleic acid fragment from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101. In certain embodiments, the nucleic acid fragment encodes a peptide that has the same activity as a peptide encoded by SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101. In certain embodiments, the activity is a kinase and/or phosphatase activity.
[0015] In certain embodiments, the transgenic plant comprises a plant kinase and/or phosphatase homologue, derivative, or variant having a nucleic acid sequence of the SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101. In certain embodiments, the homologue, derivative or variant has 1-5, 6-10, 11-20, 21-30, 31-40, 41-50, 50-70, 71-80, 81-100 nucleic acid deletion, substitution or insertion in the sequence as compared to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, or 101. In certain embodiments, the variants encode a peptide that has the same activity as a peptide encoded by SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101. In certain embodiments, the activity is a kinase and/or phosphatase activity.
[0016] In certain embodiments provided herein are the methods of altering the expression levels of plant kinase and/or phosphatase. In certain embodiments, the method comprises transforming a plant or plant cell with a nucleic acid molecule that expresses a plant kinase and/or phosphatase peptide, fragment, derivative or variant from a peptide having an amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100, or 102. In certain embodiments, the peptide, fragment, derivative or variant is overexpressed. In certain embodiments, provided herein are methods of regenerating, from said transformed plant or plant cell, a plant having enhanced growth and/or yield.
[0017] In certain embodiments, the transgenic plants express a plant kinase and/or phosphatase peptide, fragment, derivative or variant from a protein having an amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100, or 102. In certain embodiments, the peptide fragment, derivative or variant is overexpressed. In certain embodiments, provided herein are regenerated transformed plant having enhanced growth and/or yield.
4. BRIEF DESCRIPTION OF THE FIGURES
[0018] The patent application file contains at least one drawing executed in color. Copies of this patent application with color drawings will be provided by the Office upon request and payment of the necessary fee.
[0019] FIG. 1 shows a heat map of the microarray analysis of gene expression profile of Arabidopsis shoots, using three biological replicates for wild-type (WT), 2 biological replicates for AtPAP2 T-DNA line (P2), and 3 biological replicates for two independent AtPAP2 overexpression lines (OE7 and OE21).
[0020] FIG. 2 shows scatter plots of the microarray analysis of gene expression profile of Arabidopsis shoots. The results showed that the expression profiles of the two independent AtPAP2 overexpression lines (OE7 and OE21) were significantly different from that of the wild-type (WT), whereas the expression profile of the AtPAP2 T-DNA (mutant) lines resembled closely that of the WT.
[0021] FIG. 3 shows a schematic diagram of the expression vector pCXSN. (a). The cDNAs of the NG genes were cloned into the pCXSN vector at the XcmI sites to create the overexpression vectors. (b) shows an exemplified overexpression vector pCXSN-NG6.
[0022] FIG. 4 shows the mRNA expression levels of NG genes in the respective overexpression lines. The mRNA expression levels in 10-day-old T3 homologous seedlings were determined by quantitative RT-PCR using gene-specific primers. The fold-changes represent the relative expression levels of mRNAs compared to that of the wild-type (WT=1.0). The results of two trials were obtained from two batches of plant growth studies.
[0023] FIG. 5 shows the growth performance of the wild-type and NG6 over-expression lines in soil. The five columns of plants from left to right were AtPAP2 overexpression lines, WT, T3 homologous NG6 overexpression lines NG6-1, NG6-2, and NG6-3. (a) 22-day-old and (b) 25-day-old plants.
[0024] FIG. 6 shows the growth performance of the wild-type and T3 homologous NG21, NG24, NG28 and NG32 overexpression lines in soil. The five columns of plants from left to right were WT, NG21, NG24, NG28 and NG32 overexpression lines. (a) 30-day-old plants and (b) 34-day-old plants grown in black tray. (c) 22-day-old plants, (d) 25-day-old plants, (e) 28-day-old plants, and (f) 36-day-old plants grown in white cups.
4.1. BRIEF DESCRIPTION OF THE SEQUENCES
[0025] SEQ ID NO:1 is a nucleic acid sequence of Arabidopsis phosphatase NG6 gene.
[0026] SEQ ID NO:2 is an amino acid sequence of Arabidopsis phosphatase NG6.
[0027] SEQ ID NO:3 is a nucleic acid sequence of maize phosphatase NG6 gene.
[0028] SEQ ID NO:4 is an amino acid sequence of maize phosphatase NG6.
[0029] SEQ ID NO:5 is a nucleic acid sequence of soybean phosphatase NG6 gene.
[0030] SEQ ID NO:6 is an amino acid sequence of soybean phosphatase NG6.
[0031] SEQ ID NO:7 is a nucleic acid sequence of rice phosphatase NG6 gene.
[0032] SEQ ID NO:8 is an amino acid sequence of rice phosphatase NG6.
[0033] SEQ ID NO:9 is a nucleic acid sequence of cotton phosphatase NG6 gene.
[0034] SEQ ID NO:10 is an amino acid sequence of cotton phosphatase NG6.
[0035] SEQ ID NO:11 is a nucleic acid sequence of Arabidopsis kinase NG21 gene.
[0036] SEQ ID NO:12 is an amino acid sequence of Arabidopsis kinase NG21.
[0037] SEQ ID NO:13 is a nucleic acid sequence of maize kinase NG21 gene.
[0038] SEQ ID NO:14 is an amino acid sequence of maize kinase NG21.
[0039] SEQ ID NO:15 is a nucleic acid sequence of soybean kinase NG21 gene.
[0040] SEQ ID NO:16 is an amino acid sequence of soybean kinase NG21.
[0041] SEQ ID NO:17 is a nucleic acid sequence of rice kinase NG21 gene.
[0042] SEQ ID NO:18 is an amino acid sequence of rice kinase NG21.
[0043] SEQ ID NO:19 is a nucleic acid sequence of cotton kinase NG21 gene.
[0044] SEQ ID NO:20 is an amino acid sequence of cotton kinase NG21.
[0045] SEQ ID NO:21 is a nucleic acid sequence of Arabidopsis kinase NG24 gene.
[0046] SEQ ID NO:22 is an amino acid sequence of Arabidopsis kinase NG24.
[0047] SEQ ID NO:23 is a nucleic acid sequence of maize kinase NG24 gene.
[0048] SEQ ID NO:24 is an amino acid sequence of maize kinase NG24.
[0049] SEQ ID NO:25 is a nucleic acid sequence of soybean kinase NG24 gene.
[0050] SEQ ID NO:26 is an amino acid sequence of soybean kinase NG24.
[0051] SEQ ID NO:27 is a nucleic acid sequence of rice kinase NG24 gene.
[0052] SEQ ID NO:28 is an amino acid sequence of rice kinase NG24.
[0053] SEQ ID NO:29 is a nucleic acid sequence of cotton kinase NG24 gene.
[0054] SEQ ID NO:30 is an amino acid sequence of cotton kinase NG24.
[0055] SEQ ID NO:31 is a nucleic acid sequence of Arabidopsis kinase NG28 gene.
[0056] SEQ ID NO:32 is an amino acid sequence of Arabidopsis kinase NG28.
[0057] SEQ ID NO:33 is a nucleic acid sequence of maize kinase NG28 gene.
[0058] SEQ ID NO:34 is an amino acid sequence of maize kinase NG28.
[0059] SEQ ID NO:35 is a nucleic acid sequence of soybean kinase NG28 gene.
[0060] SEQ ID NO:36 is an amino acid sequence of soybean kinase NG28.
[0061] SEQ ID NO:37 is a nucleic acid sequence of rice kinase NG28 gene.
[0062] SEQ ID NO:38 is an amino acid sequence of rice kinase NG28.
[0063] SEQ ID NO:39 is a nucleic acid sequence of cotton kinase NG28 gene.
[0064] SEQ ID NO:40 is an amino acid sequence of cotton kinase NG28.
[0065] SEQ ID NO:41 is a nucleic acid sequence of Arabidopsis kinase NG32 gene.
[0066] SEQ ID NO:42 is an amino acid sequence of Arabidopsis kinase NG32.
[0067] SEQ ID NO:43 is a nucleic acid sequence of maize kinase NG32 gene.
[0068] SEQ ID NO:44 is an amino acid sequence of maize kinase NG32.
[0069] SEQ ID NO:45 is a nucleic acid sequence of soybean kinase NG32 gene.
[0070] SEQ ID NO:46 is an amino acid sequence of soybean kinase NG32.
[0071] SEQ ID NO:47 is a nucleic acid sequence of rice kinase NG32 gene.
[0072] SEQ ID NO:48 is an amino acid sequence of rice kinase NG32.
[0073] SEQ ID NO:49 is a nucleic acid sequence of cotton kinase NG32 gene.
[0074] SEQ ID NO:50 is an amino acid sequence of cotton kinase NG32.
[0075] SEQ ID NO:51 is a primer sequence useful according to the present invention.
[0076] SEQ ID NO:52 is a primer sequence useful according to the present invention.
[0077] SEQ ID NO:53 is a primer sequence useful according to the present invention.
[0078] SEQ ID NO:54 is a primer sequence useful according to the present invention.
[0079] SEQ ID NO:55 is a primer sequence useful according to the present invention.
[0080] SEQ ID NO:56 is a primer sequence useful according to the present invention.
[0081] SEQ ID NO:57 is a primer sequence useful according to the present invention.
[0082] SEQ ID NO:58 is a primer sequence useful according to the present invention.
[0083] SEQ ID NO:59 is a primer sequence useful according to the present invention.
[0084] SEQ ID NO:60 is a primer sequence useful according to the present invention.
[0085] SEQ ID NO:61 is a primer sequence useful according to the present invention.
[0086] SEQ ID NO:62 is a primer sequence useful according to the present invention.
[0087] SEQ ID NO:63 is a primer sequence useful according to the present invention.
[0088] SEQ ID NO:64 is a primer sequence useful according to the present invention.
[0089] SEQ ID NO:65 is a primer sequence useful according to the present invention.
[0090] SEQ ID NO:66 is a primer sequence useful according to the present invention.
[0091] SEQ ID NO:67 is a primer sequence useful according to the present invention.
[0092] SEQ ID NO:68 is a primer sequence useful according to the present invention.
[0093] SEQ ID NO:69 is a primer sequence useful according to the present invention.
[0094] SEQ ID NO:70 is a primer sequence useful according to the present invention.
[0095] SEQ ID NO:71 is a primer sequence useful according to the present invention.
[0096] SEQ ID NO:72 is a primer sequence useful according to the present invention.
[0097] SEQ ID NO:73 is a nucleic acid sequence of Arabidopsis AtPAP2 phosphatase gene.
[0098] SEQ ID NO:74 is an amino acid sequence of Arabidopsis AtPAP2 phosphatase.
[0099] SEQ ID NO:75 is an amino acid sequence of a conserved motif of an NG6 protein.
[0100] SEQ ID NO:76 is an amino acid sequence of a conserved motif of an NG6 protein.
[0101] SEQ ID NO:77 is an amino acid sequence of a conserved motif of an NG6 protein.
[0102] SEQ ID NO:78 is an amino acid sequence of a conserved motif of an NG6 protein.
[0103] SEQ ID NO:79 is an amino acid sequence of a conserved motif of an NG21 protein.
[0104] SEQ ID NO:80 is an amino acid sequence of a conserved motif of an NG21 protein.
[0105] SEQ ID NO:81 is an amino acid sequence of a conserved motif of an NG21 protein.
[0106] SEQ ID NO:82 is an amino acid sequence of a conserved motif of an NG21 protein.
[0107] SEQ ID NO:83 is an amino acid sequence of a conserved motif of an NG24 protein.
[0108] SEQ ID NO:84 is an amino acid sequence of a conserved motif of an NG24 protein.
[0109] SEQ ID NO:85 is an amino acid sequence of a conserved motif of an NG24 protein.
[0110] SEQ ID NO:86 is an amino acid sequence of a conserved motif of an NG28 protein.
[0111] SEQ ID NO:87 is an amino acid sequence of a conserved motif of an NG28 protein.
[0112] SEQ ID NO:88 is an amino acid sequence of a conserved motif of an NG28 protein.
[0113] SEQ ID NO:89 is an amino acid sequence of a conserved motif of an NG32 protein.
[0114] SEQ ID NO:90 is an amino acid sequence of a conserved motif of an NG32 protein.
[0115] SEQ ID NO:91 is an amino acid sequence of a conserved motif of an NG32 protein.
[0116] SEQ ID NO:92 is an amino acid sequence of a conserved motif of an NG32 protein.
[0117] SEQ ID NO:93 is a nucleic acid sequence of rapeseed kinase NG6 gene.
[0118] SEQ ID NO:94 is an amino acid sequence of rapeseed kinase NG6.
[0119] SEQ ID NO:95 is a nucleic acid sequence of rapeseed kinase NG21 gene.
[0120] SEQ ID NO:96 is an amino acid sequence of rapeseed kinase NG21.
[0121] SEQ ID NO:97 is a nucleic acid sequence of rapeseed kinase NG24 gene.
[0122] SEQ ID NO:98 is an amino acid sequence of rapeseed kinase NG24.
[0123] SEQ ID NO:99 is a nucleic acid sequence of rapeseed kinase NG28 gene.
[0124] SEQ ID NO:100 is an amino acid sequence of rapeseed kinase NG28.
[0125] SEQ ID NO:101 is a nucleic acid sequence of rapeseed kinase NG32 gene.
[0126] SEQ ID NO:102 is an amino acid sequence of rapeseed kinase NG32.
5. DETAILED DESCRIPTION
[0127] Provided herein are methods of producing plants with enhanced growth and/or yield. In one embodiment, the method comprises: transforming a plant or plant cell with a nucleic acid molecule comprising a plant kinase and/or phosphatase gene selected from NG6, NG21, NG24, NG28, and NG32, and over-expressing said kinase and/or phosphatase gene in the plant or plant cell. In one embodiment, the method further comprises: regenerating, from said transformed plant or plant cell, a plant having enhanced growth and/or yield. Also provided are transgenic plants with enhanced growth and/or yield, comprising a plant kinase and/or phosphatase gene selected from NG6, NG21, NG24, NG28, and NG32, wherein the kinase and/or phosphatase is overexpressed in the plant or plant cell.
[0128] The inventors discover that altering the expression levels of one or more phosphatases (such as NG6) and kinases (such as NG21, NG24, NG28, and NG32) results in rapid plant growth and higher yield. The gene expression profiles of the AtPAP2 overexpression lines, AtPAP2 T-DNA (mutant) line, and the wild-type plants are analyzed using microarray. The microarray data show that the expression levels of a range of genes are significantly altered (upregulated or downregulated) in the AtPAP2 overexpression lines, when compared to the wild-type.
[0129] The introduction of a representative gene of phosphatases (AT1G05000 (NG6)) and kinases (AT1G13350 (NG21), AT1G28390 (NG24), AT3G24660 (NG28) and AT5G03320 (NG32)), into the genome of Arabidopsis by transgenic technology produced transgenic Arabidopsis that grew faster than the wild-type plants (Table 4, FIG. 5, FIG. 6), and the yield of seeds were elevated by 23-70% (Table 5).
[0130] While any plant species can be modified using the methods described herein, preferably included without limitation are species from the following genera with representative species in parentheses:
[0131] Monocots: genera Asparagus (asparagus), Bromus (cheatgrass), Hemerocallis (daylily), Hordeum (barley), Lolium (ryegrass), Oryza (rice), Panicum (Switchgrass), Pennisetum (fountaingrass), Saccharum (Sugar cane), Sorghum, Trigonella (fenu grass), Triticum (wheat), and Zea (corn); and
[0132] Dicots: genera Antirrhinum (flower sp.), Arabidopsis (thaliana), Arachis (peanut), Atropa (deadly nightshade), Brassica (rapeseed), Browallia, Capsicum (pepper), Carthamus (safflower), Cichorium (chicory), Citrus (orange, lemon), Chrysanthemum, Cucumis (cucumber), Datura (thorn apple), Daucus (carrot), Digitalis (foxglove), Fragaria (strawberry), Geranium (flower sp.), Glycine (soybean), Helianthus (sunflower), Hyscyamus, Ipomoea (morning glory), Latuca (lettuce), Linum (linseed), Lotus (flower sp.), Lycopersicon (tomato), Majorana, Malva (cotton), Manihot, Medicago (alfalfa), Nemesia, Nicotiana (tobacco), Onobrychis, Pelargonium (citrosa), Petunia (flower sp.), Ranunculus (flower sp.), Raphanus (radishes), Salpiglossis, Senecio (flower sp.), Sinapis (albae semen), Solanum (potato), Trifolium (clovers), Vigna (mungbean, fava bean), and Vitis (grape).
[0133] In certain embodiments, plant species transgenically modified according to the present invention are selected from soybean, maize, potato, rice, sugar canes, switchgrass, cotton, sorghum, alfalfas, rapeseed, canola, rye, sorghum, sunflower, wheat, tobacco, millet, peanuts sweet potato cassava, coffee, coconut, cocoa, tea, banana, citrus, apple, pineapple, avocado, fig, guava, mango, olive, barley ornamentals, and conifers. In preferred embodiments, plant species transgenically modified according to the present invention are selected from soybean, maize, potato, rice, sugar canes, switchgrass, cotton, sorghum, alfalfas, rapeseed, and canola.
[0134] In certain embodiment, plant parts, plant tissue, and plant cells including, but not limited to, shoots, stems, seeds, and roots, can be transgenically modified in accordance with the present invention.
4.2 Definitions
[0135] The term "protein or peptide homologue," as used herein, refers to one or more of the following proteins or peptides: (i) a protein or polypeptide with at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 98% sequence identity with a protein or polypeptide of the invention; (ii) a protein or polypeptide encoded by a nucleotide sequence that is at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 98% identical to a nucleic acid sequence of the invention; (iii) a protein or polypeptide encoded by a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence of the invention; (iv) a protein or polypeptide that is derived from conservative substitution of amino acids of a protein or polypeptide of the invention, or that is derived from conservative substitution of amino acids of a protein or polypeptide of (i)-(iii); (v) a fragment of a protein or polypeptide of the invention or a fragment of a protein or polypeptide of (i) through (iv); and (vii) a protein or polypeptide recognized by an antibody that immunospecifically binds to a sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102.
[0136] The term "an antibody or an antibody fragment that immunospecifically binds to a polypeptide selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102" or "an antibody or an antibody fragment that immunospecifically binds to a polypeptide, peptide, or protein of the invention," as used herein, refers to an antibody or a fragment thereof that immunospecifically binds to a polypeptide selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102, or a fragment of these polypeptide, wherein the antibody or the antibody fragment does not non-specifically bind to other peptides, polypeptides, or proteins.
[0137] An antibody or a fragment thereof that immunospecifically binds to a polypeptide selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102, or a fragment of these polypeptide, may cross-react with other antigens. In a preferred embodiment, an antibody or a fragment thereof that immunospecifically binds to a polypeptide selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102, or a fragment of these polypeptides, does not cross-react with other antigens. An antibody or a fragment thereof that immunospecifically binds to a polypeptide selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102, or a fragment of these polypeptide, can be identified by, for example, immunoassays or other techniques known to those skilled in the art. An antibody or an antibody fragment that immunospecifically binds a polypeptide selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102 may be interchangeably referred to as "anti-PAP antibody".
[0138] The term "peptide or protein derivative," as used herein, refers to a given peptide or protein that is modified, e.g., by covalent attachment of another molecule, to the peptide or protein, including the incorporation of non-naturally occurring amino acids. The peptide or protein derivative retains one or more biological activities of the peptide or protein.
[0139] The term "nucleic acid fragment," as used herein, refers to a fragment of a nucleic acid molecule of the invention, wherein the fragment comprises at least about 400, at least about 450, at least about 500, at least about 550, at least about 600, at least about 650, at least about 700, at least about 750, at least about 800, at least about 850, at least about 900, at least about 950, at least about 1000, at least about 1050, at least about 1100, at least about 1150, at least about 1200, at least about 1250, at least about 1300, or at least about 1350 contiguous nucleic acid bases of the nucleic acid molecule.
[0140] The term "protein or peptide fragment," as used herein, refers to a fragment of a protein or peptide of the invention, wherein the fragment comprises at least about 160, at least about 180, at least about 200, at least about 220, at least about 240, at least about 260, at least about 280, at least about 300, at least about 320, at least about 340, or at least about 360 contiguous amino acid residues of the protein or peptide.
[0141] The term "protein or peptide variant," as used herein, includes 1) a naturally occurring allelic variation of a given protein or peptide, and 2) a recombinantly prepared variation of a given protein or peptide, in which one or more amino acid residues have been modified by amino acid substitution, addition, and/or deletion.
[0142] An "isolated" nucleic acid molecule has been removed from any environment in which it may exist in nature. For instance, an "isolated" nucleic acid molecule, such as a cDNA molecule, is substantially free of other cellular materials, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. In a preferred embodiment, nucleic acid molecules encoding the polypeptides/proteins of the present invention are isolated or purified.
[0143] The term "under stringent conditions" refers to hybridization and washing conditions under which nucleotide sequences having homology to each other remain hybridized to each other. Such hybridization conditions are described in, for example, but not limited to, Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6.; Basic Methods in Molecular Biology, Elsevier Science Publishing Co., Inc., N.Y. (1986), pp. 75-78, and 84-87; and Molecular Cloning, Cold Spring Harbor Laboratory, N.Y. (1982), pp. 387-389, and are well known to those skilled in the art. A preferred example of stringent hybridization conditions is hybridization in 6× sodium chloride/sodium citrate (SSC), 0.5% SDS at about 68° C. followed by one or more washes in 2×SSC, 0.5% SDS at room temperature. Another preferred, example of stringent hybridization conditions is hybridization in 6×SSC at about 45° C. followed by one or more washes in 0.2×SSC, 0.1% SDS at about 50-65° C.
[0144] To determine the percent identity of two amino acid sequences or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=number of identical overlapping positions/total number of positions×100%). In one embodiment, the two sequences are the same length.
[0145] The determination of percent identity between two sequences can also be accomplished using a mathematical algorithm. A preferred, non limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. U.S.A. 87:2264 2268, modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. U.S.A. 90:5873 5877. Such an algorithm is incorporated into the NBLAST and) (BLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecules of the present invention. BLAST protein searches can be performed with the XBLAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule of the present invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389 3402. Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., the NCBI website). Another preferred, non limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11 17. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.
[0146] The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.
[0147] As used herein, the term "derivative" (e.g., proteins, polypeptides, peptides, and antibodies) refers to an agent that comprises an amino acid sequence which has been altered by the introduction of amino acid residue substitutions, deletions, and/or additions. The term "derivative" as used herein also refers to an agent which has been modified, i.e., by the covalent attachment of any type of molecule to the agent. For example, but not by way of limitation, an antibody may be modified, e.g., by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. A derivative of an agent may be produced by chemical modifications using techniques known to those of skill in the art, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Further, a derivative of an agent may contain one or more non-classical amino acids. A derivative of an agent possesses a similar or identical function as the agent from which it was derived.
[0148] The term "enhance or promote plant growth and/or yield" refers to for example, increased plant weight, increased leaf number and/or weight, increased number of inflorescence, increased seed production (such as weight/seed and total weight of seeds), increased carbon metabolism, increased carbohydrate (e.g., starch, sugars, cellulose), amino acid, and/or lipid production, early bolting, and also can include combinations of the foregoing, when compared to a wild-type plant of the same species cultivated under the same conditions.
5.1 Growth-promoting Phosphatases and Kinases
[0149] Provided herein are phosphatases and kinases that promote plant growth and/or yield. In one embodiment, the growth-promoting phosphatase is NG6, and the growth-promoting kinases are selected from NG6, NG21, NG24, NG28, and NG32. In certain specific embodiments, the growth-promoting phosphatases and kinases are derived from plant species including, but not limited to, Arabidopsis, rice, soybean, maize, and cotton.
[0150] In certain embodiments, the phosphatase gene that promotes plant growth and/or yield is an NG6 gene comprising a nucleic acid sequence selected from SEQ ID NO: 1, 3, 5, 7, 9 or 93. In certain embodiments, the phosphatase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 1, 3, 5, 7, 9 or 93. In certain embodiments, the phosphatase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule derives from a nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 1, 3, 5, 7, 9 or 93.
[0151] In certain embodiments, the phosphatase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 1, 3, 5, 7, 9 or 93. In certain embodiments, the phosphatase gene that promotes plant growth and/or yield comprises the nucleic acid sequence that encodes a protein that comprises one or more of the following conserved motifs: GIFRSGFP (SEQ ID NO:75), YLCPEPYP (SEQ ID NO:76), KEPFVXIP (SEQ ID NO:77), and HCXRGKHRTG (SEQ ID NO:78).
[0152] In certain embodiments, the phosphatase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequences comprising SEQ ID NO: 1, 3, 5, 7, 9 or 93. In certain embodiments, the phosphatase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: GIFRSGFP (SEQ ID NO:75), YLCPEPYP (SEQ ID NO:76), KEPFVXIP (SEQ ID NO:77), and HCXRGKHRTG (SEQ ID NO:78).
[0153] In certain embodiments, the kinase gene that promotes plant growth and/or yield is an NG21 gene comprising a nucleic acid sequence selected from SEQ ID NO: 11, 13, 15, 17, 19 or 95. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 11, 13, 15, 17, 19 or 95. In certain embodiments, the kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 11, 13, 15, 17, 19 or 95.
[0154] In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 11, 13, 15, 17, or 19 or 95. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises the nucleic acid sequence that encodes a protein that comprises one or more of the following conserved motifs: DNWDDA(D/E)GYY (SEQ ID NO:79), YRNHLCLVFESL (SEQ ID NO:80), VLHCDIKPDNMLVNE (SEQ ID NO:81), and TPYLVSRFYRXPEI (SEQ ID NO:82).
[0155] In certain embodiments, the kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequences comprising SEQ ID NO: 11, 13, 15, 17, 19 or 95. In ceratin embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: DNWDDA(D/E)GYY (SEQ ID NO:79), YRNHLCLVFESL(SEQ ID NO:80), VLHCDIKPDNMLVNE (SEQ ID NO:81), and TPYLVSRFYRXPEI (SEQ ID NO:82). In certain embodiments, the kinase gene that promotes plant growth and/or yield is an NG24 gene comprising a nucleic acid sequence selected from SEQ ID NO: 21, 23, 25, 27, 29 or 97. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 21, 23, 25, 27, 29 or 97. In certain embodiments, the kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 21, 23, 25, 27, 29 or 97.
[0156] In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 21, 23, 25, 27, 29 or 97. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: VRHRDXKS (SEQ ID NO:83), GTLXGYLDP (SEQ ID NO:84), and DV(F/Y)S(F/Y)G(I/V)LLLEI (SEQ ID NO:85).
[0157] In certain embodiments, the kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 21, 23, 25, 27, 29 or 97. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: VRHRDXKS (SEQ ID NO:83), GTLXGYLDP (SEQ ID NO:84), and DV(F/Y)S(F/Y)G(I/V)LLLEI (SEQ ID NO:85). In certain embodiments, the kinase gene that promotes plant growth and/or yield is an NG28 gene comprising a nucleic acid sequence selected from SEQ ID NO: 31, 33, 35, 37, 39 or 99. In certain embodiments, the NG24 kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 31, 33, 35, 37, 39 or 99. In certain embodiments, the NG24 kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 31, 33, 35, 37, 39 or 99.
[0158] In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 31, 33, 35, 37, 39 or 99. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: RRHKIALG (SEQ ID NO:86), Y(K/R)APEL (SEQ ID NO:87), and DVYAFGILLLE (SEQ ID NO:88).
[0159] In certain embodiments, the kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 31, 33, 35, 37, 39 or 99. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: RRHKIALG (SEQ ID NO:86), Y(K/R)APEL (SEQ ID NO:87), and DVYAFGILLLE (SEQ ID NO:88).
[0160] In certain embodiments, the kinase gene that promotes plant growth and/or yield is an NG32 gene comprising a nucleic acid sequence selected from SEQ ID NO: 41, 43, 45, 47, 49 or 101. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 41, 43, 45, 47, 49 or 101. In certain embodiments, the kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 41, 43, 45, 47, 49 or 101.
[0161] In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises a nucleic acid sequence having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 41, 43, 45, 47, 49 or 101. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: CAXDDERG (SEQ ID NO:89), AKLSDFGLAR (SEQ ID NO:90), YELITGR(R/K) (SEQ ID NO:91), and RPKMSEV (SEQ ID NO:92).
[0162] In certain embodiments, the kinase gene that promotes plant growth and/or yield is a homologue, derivative, or variant of a nucleic acid molecule that derives from nucleic acid molecule having nucleic acid sequence comprising SEQ ID NO: 41, 43, 45, 47, 49 or 101. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: CAXDDERG (SEQ ID NO:89), AKLSDFGLAR (SEQ ID NO:90), YELITGR(R/K) (SEQ ID NO:91), and RPKMSEV (SEQ ID NO:92). In certain embodiments, the phosphatase or kinase gene that promotes plant growth and/or yield encodes a protein selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the phosphatase or kinase gene that promotes plant growth and/or yield encodes a protein having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the phosphatase or kinase gene that promotes plant growth and/or yield encodes a protein that is a homologue, derivative, or variant of a protein derived from the amino acid molecule having the amino acid sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102.
[0163] In certain embodiments, the phosphatase gene that promotes plant growth and/or yield encodes an NG6 protein having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 2, 4, 6, 8, 10 or 94. In certain embodiments, the phosphatase gene that promotes plant growth and/or yield one or more of the following conserved motifs: GIFRSGFP (SEQ ID NO:75), YLCPEPYP (SEQ ID NO:76), KEPFVXIP (SEQ ID NO:77), and HCXRGKHRTG (SEQ ID NO:78).
[0164] In certain embodiments, the phosphatase gene that promotes plant growth and/or yield encodes an NG6 protein that is a homologue, derivative, or variant of a protein derived from the amino acid molecule having the amino acid sequence comprising SEQ ID NO: 2, 4, 6, 8, 10 or 94. In certain embodiments, the phosphatase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: GIFRSGFP (SEQ ID NO:75), YLCPEPYP (SEQ ID NO:76), KEPFVXIP (SEQ ID NO:77), and HCXRGKHRTG (SEQ ID NO:78).
[0165] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG 21 protein having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 12, 14, 16, 18, 20 or 96. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: DNWDDA(D/E)GYY (SEQ ID NO:79), YRNHLCLVFESL (SEQ ID NO:80), VLHCDIKPDNMLVNE (SEQ ID NO:81), and TPYLVSRFYRXPEI (SEQ ID NO:82).
[0166] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG21 protein that is a homologue, derivative, or variant of a protein derived from the amino acid molecule having the amino acid sequence selected from SEQ ID NO: 12, 14, 16, 18, 20 or 96. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: DNWDDA(D/E)GYY (SEQ ID NO:79), YRNHLCLVFESL (SEQ ID NO:80), VLHCDIKPDNMLVNE (SEQ ID NO:81), and TPYLVSRFYRXPEI (SEQ ID NO:82).
[0167] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG24 protein having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 22, 24, 26, 28, 30 or 98. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: VRHRDXKS (SEQ ID NO:83), GTLXGYLDP (SEQ ID NO:84), and DV(F/Y)S(F/Y)G(I/V)LLLEI (SEQ ID NO:85).
[0168] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG24 protein that is a homologue, derivative, or variant of a protein derived from the amino acid molecule having the amino acid sequence selected from SEQ ID NO: 22, 24, 26, 28, 30 or 98. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: VRHRDXKS (SEQ ID NO:83), GTLXGYLDP (SEQ ID NO:84), and DV(F/Y)S(F/Y)G(UV)LLLEI (SEQ ID NO:85).
[0169] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG28 protein having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 32, 34, 36, 38, 40 or 100. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: RRHKIALG (SEQ ID NO:86), Y(K/R)APEL (SEQ ID NO:87), and DVYAFGILLLE (SEQ ID NO:88).
[0170] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG28 protein that is a homologue, derivative, or variant of a protein derived from the amino acid molecule having the amino acid sequence selected from SEQ ID NO: 32, 34, 36, 38, 40 or 100, wherein the protein comprises one or more of the following conserved motifs: RRHKIALG (SEQ ID NO:86), Y(K/R)APEL (SEQ ID NO:87), and DVYAFGILLLE (SEQ ID NO:88).
[0171] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG32 protein having at least 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 42, 44, 46, 48, 50 or 102. In certain embodiments, the kinase gene that promotes plant growth and/or yield comprises one or more of the following conserved motifs: CAXDDERG (SEQ ID NO:89), AKLSDFGLAR (SEQ ID NO:90), YELITGR(R/K) (SEQ ID NO:91), and RPKMSEV (SEQ ID NO:92).
[0172] In certain embodiments, the kinase gene that promotes plant growth and/or yield encodes an NG32 protein that is a homologue, derivative, or variant of a protein derived from the amino acid molecule having the amino acid sequence selected from SEQ ID NO: 42, 44, 46, 48, 50 or 102, wherein the protein comprises one or more of the following conserved motifs: CAXDDERG (SEQ ID NO:89), AKLSDFGLAR (SEQ ID NO:90), YELITGR(R/K) (SEQ ID NO:91), and RPKMSEV (SEQ ID NO:92).
5.2 Production of Transgenic Plants with Enhanced Growth and/or Yield
[0173] Another aspect of the present invention provides methods of producing plants with enhanced growth and/or yield. In one embodiment, the method comprises: transforming a plant or plant cell with a nucleic acid molecule comprising a plant kinase and/or phosphatase gene of the present invention. In one embodiment, the method comprises overexpressing said kinase and/or phosphatase gene in the plant or plant cell. In one embodiment, the present invention further comprises: regenerating, from said transformed plant or plant cell, a plant having enhanced growth and/or yield.
[0174] The term "overexpressing," "overexpression," or any of the grammatical variations thereof (e.g., over-expressing, over-expression) refers to an increase in the level of expression of a gene, or the level of a protein product encoded by a gene, wherein such increase is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200%, when compared to cells of the same type in a wild-type plant of the same species cultivated under the same conditions.
[0175] In one embodiment, the method further comprises: transforming a plant or a plant cell with a nucleic acid molecule comprising an AtPAP2 gene. In certain embodiments, the method comprises overexpressing the AtPAP2 gene in the plant or plant cell. In one embodiment, the AtPAP2 gene comprises SEQ ID NO: 73. In certain embodiments, the AtPAP2 gene comprises a nucleic acid molecule having a nucleic acid molecule having sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 73.
[0176] In one embodiment, the method further comprises: transforming a plant or a plant cell with a nucleic acid molecule encoding AtPAP2 phosphatase. In certain embodiment, the method comprises overexpressing the nucleic acid molecule encoding AtPAP2 phosphatase in the plant or plant cell. In one embodiment, AtPAP2 phosphatase comprises SEQ ID NO: 74. In certain embodiments, AtPAP2 phosphatase comprises an amino acid molecule having an amino acid nucleic acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 74.
[0177] Provided herein are transgenic plants with enhanced growth and/or yield. In certain embodiments, the transgenic plant comprises a nucleic acid molecule having a nucleic acid sequence selected from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101. In certain embodiments, the nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions. In certain embodiments, the transgenic plant comprises a nucleic acid molecule having a nucleic acid sequence that is at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101. In certain embodiments, the nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions. In certain embodiments, the transgenic plant comprises a nucleic acid molecule that is a homologue, derivative, or variant of a nucleic acid molecule derived from the nucleic acid molecule having a nucleic acid sequence selected from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101. In certain embodiments, the nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions.
[0178] In certain embodiments, the transgenic plant comprises a nucleic acid that encodes a protein having an amino acid sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions. In certain embodiments, the transgenic plant comprises a nucleic acid that encodes a protein having an amino acid sequence that is at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions. In certain embodiments, the transgenic plant comprises a nucleic acid that encodes a protein that is a homologue, derivative, or variant of a protein derived from the peptide having an amino acid sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the nucleic acid molecule is overexpressed in the transgenic plant when compared to a wild-type plant of the same species cultivated under the same conditions.
[0179] In certain embodiments, the transgenic plant comprises a protein having an amino acid sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the level of the protein in the transgenic plant is higher than that of a wild-type plant of the same species cultivated under the same conditions. In certain embodiments, the transgenic plant comprises a protein having an amino acid sequence that is at least 65%, 70%, 75%, 80%, 85%, 90%, 93%, 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the level of the protein in the transgenic plant is higher than that of a wild-type plant of the same species cultivated under the same conditions. In certain embodiments, the transgenic plant comprises a protein that is a homologue, derivative, or variant of a protein derived from the peptide having an amino acid and sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 94, 96, 98, 100 or 102. In certain embodiments, the level of the protein in the transgenic plant is higher than that of a wild-type plant of the same species cultivated under the same conditions.
[0180] In addition, the present invention provides transgenic plant cells transformed with a nucleic acid molecule of the present invention. In one embodiment, the invention provides transgenic plant cells comprising a kinase or phosphatase nucleic acid molecule of the invention. In certain embodiments, the nucleic acid molecule is overexpressed in the transgenic plant cells when compared to plant cells of the same type in a wild-type plant of the same species cultivated under the same conditions. In another embodiment, the invention provides transgenic plant cells comprising a kinase or phosphatase protein of the invention, wherein the level of said protein in the transgenic plant cells is higher (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher) than that of plant cells of the same type in a wild-type plant of the same species cultivated under the same conditions.
[0181] In certain embodiments, the transgenic plant comprises a nucleic acid molecule encoding a phosphatase having an amino acid sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, or 94 and/or a kinase selected from SEQ ID NO: 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 96, 98, 100 or 102. In another embodiment, the transgenic plant comprises a nucleic acid molecule encoding a phosphatase having an amino acid sequence selected from SEQ ID NO: 2, 4, 6, 8, 10, or 94 and/or a kinase selected from SEQ ID NO: 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 96, 98, 100 or 102. In certain embodiment, all or a portion, particularly an N-terminal portion, of amino acid residues 1 to 80, preferably all or a portion of amino acid residues 1 to 30, are replaced by a heterologous plant signal peptide by genetic engineering. In such a transgenic plant, the phosphatases or kinases are directed to various organelles/compartments of the cells.
[0182] In certain embodiments, the present invention provides chimeric gene constructs for genetic modification of plants to increase growth rate and to improve yield. In a specific embodiment, the chimeric gene constructs comprise a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101. In another specific embodiment, the chimeric gene constructs comprise a sequence that hybridizes under stringent conditions to a nucleic acid molecule comprising a nucleic acid sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101, or a complement thereof, wherein the nucleic acid sequence encodes a protein or a polypeptide that exhibits at least one structural and/or functional feature of the polypeptides and enhances plant growth and/or yield.
[0183] The phosphatase or kinase-coding sequence is operatively linked to upstream and downstream regulatory components, preferably heterologous to the phosphatase or kinase sequence, such as for example, CMV 35S promoter, which acts to cause expression of the gene (production of the enzyme) in plant cells (see FIG. 3). Preferably, when a construct comprising a gene encoding a phosphatase or kinase of the present invention is introduced into plant cells by a conventional transformation method, such as microparticle bombardment, Agrobacterium infection, or microinjection, the gene is expressed in the cells under the control of the regulatory sequences. The expressed phosphatase interacts with the biosynthetic machinery that is naturally present in the plant cells to alter the carbon metabolism. By altering the carbon metabolism, the method of the present invention promotes the growth rate of the plant, resulting in faster growth rate and higher yield. As a result, the time required for the maturation of the plant and the time required for flowering is shortened. Also provided are methods for increasing growth rate and yield of plants, comprising the step of inserting into such plant cells, or cells of such whole plants, a chimeric gene construct.
[0184] In one specific embodiment, Arabidopsis is genetically modified by introducing an overexpression construct comprising nucleic acid molecules encoding a growth-promoting phosphatase or kinase of the present invention.
[0185] In an embodiment, the growth-promoting phosphatase and kinase genes are derived from Arabidopsis. As shown in the examples, transgenic Arabidopsis plants with over-expression of NG6, NG21, NG24, NG28, and/or NG32 have enhanced growth and/or yield, when compared to wild-type Arabidopsis plants (see Table 5, and FIGS. 5 and 6).
[0186] In one embodiment, a transgenic plant overexpressing a nucleic acid comprises the sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101 or homologues thereof, wherein the nucleic acid molecule encodes polypeptides or proteins of the invention.
5.3 Homologues, Derivatives, and Variants of Kinases and Phosphatases
[0187] The present invention also provides homologues, derivatives, and variants of kinases and phosphatases of the present invention; nucleic acid molecules encoding the polypeptides and homologues, derivatives, and variants; vectors, plant cells and transgenic plants comprising these nucleic acid molecules; and uses thereof for promoting plant growth and/or yield. The homologues, derivatives and variants of kinases and phosphatases are derived from the wild-type kinases and phosphatases, respectively. The methods of deriving the homologues, derivatives and variants are well known in the art which include routine conventional techniques of chemical modifications of amino acid residues or using molecular biology and recombinant DNA manipulation and production. Such techniques are available to the skilled artisan in laboratory manuals such as Sambrook and Russell, Molecular cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2001).
[0188] In one embodiment, a homologue of the nucleic acid or polypeptide molecule of the present invention includes: (i) a polypeptide with at least about 65%, at least about 70%, at least about 80%, at least about 90%, or at least about 98% sequence identity of the polypeptide of the invention; (ii) a polypeptide encoded by a nucleotide sequence that is at least about 65%, at least about 70%, at least about 80%, at least about 90%, or at least about 98% identical to one or more of the nucleotide sequences encoding a polypeptide of the invention, or a fragment thereof; (iii) a polypeptide encoded by a nucleotide sequence that hybridizes, under stringent conditions, to a nucleotide sequence of the present invention; (iv) a polypeptide having an amino acid sequence that is at least about 65%, at least about 70%, at least about 80%, at least about 90%, or at least about 98% identical to a polypeptide of the present invention, and wherein the polypeptide of the invention is conservatively substituted; (v) a nucleic acid sequence encoding an amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, or at least about 98% identical to a polypeptide of the present invention and wherein the polypeptide of the invention is conservatively substituted; and (vi) a fragment of a polypeptide described in (i) through (iv), wherein the polypeptide fragment has at least 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 450, 500, 550, 600, 650, 700, or 750 contiguous amino acid residues of a polypeptide of the invention.
[0189] In one embodiment, a homologue polypeptide has an amino acid sequence that is at least about 65%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 98% identical to a kinase or phosphatase of the present invention. In one embodiment, the homologue polypeptide is obtained by conservative substitution.
[0190] In one aspect, the homologues derivatives and variants are derived from the wild type kinase and phosphatase by substitution, deletion, insertion of one or more nucleic acid in a nucleic acid molecule or one or more amino acid residues in an amino acid molecule. The term "derived" as used herein includes the modifications of a wild type nucleic acid molecule or amino acid molecule as described below. For example, non-natural amino acids can be substituted for the amino acids of the kinases and phosphatases so long as the kinases and phosphatases having the substituted amino acids retain substantially the same functional activity as the kinases and phosphatases in which amino acids have not been substituted. Those having skill in the art will recognize that mutations can be made to polynucleotides encoding protein and peptides, or complementary thereto, and that such mutations do not cause structural changes that affect functionality.
[0191] Conservative substitutions whereby a modified protein or polypeptide of the present invention having an amino acid of one class is replaced with another amino acid of the same class fall within the scope of the subject invention so long as the modified protein or polypeptide having the substitution still retains substantially the same functional activity as the protein or polypeptide that does not have the substitution. For instance, amino acid residue of any of the following 11 groups may be conservatively substituted with another amino acid of the same group: (1) acidic (negatively charged) amino acids, such as aspartic acid and glutamic acid; (2) basic (positively charged) amino acids, such as arginine, histidine, and lysine; (3) neutral polar amino acids, such as glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; (4) neutral nonpolar (hydrophobic) amino acids, such as alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; (5) amino acids having aliphatic side chains, such as glycine, alanine, valine, leucine, and isoleucine; (6) amino acids having aliphatic-hydroxyl side chains, such as serine and threonine; (7) amino acids having amide-containing side chains, such as asparagine and glutamine; (8) amino acids having aromatic side chains, such as phenylalanine, tyrosine, and tryptophan; (9) amino acids having basic side chains, such as lysine, arginine, and histidine; (10) amino acids having sulfur-containing side chains, such as cysteine and methionine; and (11) amino acids having similar geometry and hydrogen bonding patterns, such as aspartic acid, asparagine, glutamic acid and glutamine.
[0192] Examples of non-natural amino acids include, but are not limited to, ornithine, citrulline, hydroxyproline, homoserine, phenylglycine, taurine, iodotyrosine, 2,4-diaminobutyric acid, α-amino isobutyric acid, 4-aminobutyric acid, 2-amino butyric acid, γ-amino butyric acid, ε-amino hexanoic acid, 6-amino hexanoic acid, 2-amino isobutyric acid, 3-amino propionic acid, norleucine, norvaline, sarcosine, homocitrulline, cysteic acid, τ-butylglycine, τ-butylalanine, phenylglycine, cyclohexylalanine, β-alanine, fluoro-amino acids, designer amino acids such as β-methyl amino acids, C-methyl amino acids, N-methyl amino acids, and amino acid analogues in general. Non-natural amino acids also include amino acids having derivatized side groups. Furthermore, any of the amino acids in the protein can be of the D (dextrorotary) form or L (levorotary) form.
[0193] The structure of a polypeptide can be determined by methods known to those skilled in the art, including but not limited to, X-ray crystallography, nuclear magnetic resonance, and crystallographic electron microscopy. A sequence having sequence homology can be made using standard molecular biology techniques, including site-directed mutagenesis and by insertion or deletion of sequences.
[0194] In one aspect, the homologues, derivatives and variants are derived from the wild type kinase and phosphatase. In certain embodiments, provided herein are derivatives of the disclosed polypeptides. For example, but not by way of limitation, derivatives may include peptides or proteins that have been modified, e.g., by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. Any of numerous chemical modifications may be carried out by known techniques including, but not limited to, specific chemical cleavage, acetylation, formylation, etc. Additionally, the derivative may contain one or more non-classical amino acids. The subject invention also concerns variants of the polynucleotides of the present invention. Variant sequences include those sequences wherein one or more nucleotides of the sequence have been substituted, deleted, and/or inserted.
[0195] The nucleotides that can be substituted for natural nucleotides of DNA have a base moiety that can include, but is not limited to, inosine, 5-fluorouracil, 5-bromouracil, hypoxanthine, 1-methylguanine, 5-methylcytosine, and tritylated bases. The sugar moiety of the nucleotide in a sequence can also be modified and includes, but is not limited to, arabinose, xylulose, and hexose. In addition, the adenine, cytosine, guanine, thymine, and uracil bases of the nucleotides can be modified with acetyl, methyl, and/or thio groups. Sequences containing nucleotide substitutions, deletions, and/or insertions can be prepared and tested using standard techniques known in the art.
[0196] Unless otherwise specified, as used herein percent sequence identity and/or similarity of two sequences can be determined using the algorithm of Karlin and Altschul (1990), modified as in Karlin and Altschul (1993). Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al. (1990). BLAST searches can be performed with the NBLAST program, score=100, wordlength=12, to obtain sequences with the desired percent sequence identity. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used as described in Altschul et al. (1997). When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (NBLAST and XBLAST) can be used. See NCBI/NIH website.
[0197] The subject invention also contemplates those polynucleotide molecules having sequences which are sufficiently homologous with the polynucleotide sequences exemplified herein so as to permit hybridization with that sequence under standard stringent conditions and standard methods (Maniatis et al., 1982).
[0198] In one embodiment, the present invention further provides isolated nucleic acid molecules that comprise, or consist of, at least about 550, at least about 600, at least about 650, at least about 700, at least about 750, at least about 800, at least about 850, at least about 900, at least about 950, at least about 1000, at least about 1050, at least about 1100, at least about 1150, at least about 1200, at least about 1250, at least about 1300, or at least about 1350 contiguous nucleotides of a nucleic acid molecule of the present invention.
[0199] In another embodiment, an isolated nucleic acid molecule encodes a variant of a polypeptide whose amino acid sequence has been modified by genetic engineering so that biological activities of the polypeptides are either enhanced or reduced, or the local structures thereof are changed without significantly altering the biological activities. Amino acid modifications can be made by methods known in the art.
[0200] In one embodiment, the present invention embodies isolated nucleic acid molecules that hybridize, under stringent conditions, to nucleic acid molecules having the nucleic acid sequence comprising SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99 or 101, or homologues thereof. In certain embodiments, the nucleic acid molecules encode proteins or polypeptides that exhibit at least one structural and/or functional feature of the polypeptides of the invention (e.g. enhance plant growth and/or yield).
[0201] A further embodiment includes methods for preparing a polypeptide as provided herein by recombinant DNA technology. In one embodiment, the preparation method comprises culturing host cells containing a recombinant expression vector encoding a polypeptide as provided herein, or a nucleotide sequence encoding a polypeptide as provided herein operably linked to a heterologous promoter, and producing the polypeptide as provided herein.
5.4 Vectors and Expression Constructs
[0202] Another embodiment includes nucleic acid molecules suitable for use as primers or hybridization probes for the detection of nucleic acids encoding a phosphatase or kinase polypeptide as provided herein or other sequences.
[0203] Yet another embodiment includes vectors, e.g., recombinant expression vectors, comprising a nucleic acid molecule as provided herein. Furthermore, host cells containing such a vector or engineered to contain and/or express a nucleic acid molecule as provided herein and host cells containing a nucleotide sequence as provided herein operably linked to a heterologous promoter are disclosed.
[0204] As used herein, the term "expression construct" refers to a combination of nucleic acid sequences that provides for transcription of an operably linked nucleic acid sequence. In general, operably linked components are in contiguous relation.
[0205] Expression constructs of the invention will also generally include regulatory elements that are functional in the intended host cell in which the expression construct is to be expressed. Regulatory elements include promoters, transcription termination sequences, translation termination sequences, enhancers, and polyadenylation elements.
[0206] An expression construct as provided herein can comprise a promoter sequence operably linked to a polynucleotide sequence encoding a peptide. Promoters can be incorporated into a polynucleotide using standard techniques known in the art. Multiple copies of promoters or multiple promoters can be used in an expression construct. In a preferred embodiment, a promoter can be positioned about the same distance from the transcription start site as it is from the transcription start site in its natural genetic environment. Some variation in this distance is permitted without substantial decrease in promoter activity. A transcription start site is typically included in the expression construct.
[0207] Unique restriction enzyme sites can be included at the 5' and 3' ends of the expression construct to allow for insertion into a polynucleotide vector. As used herein, the term "vector" refers to any genetic element, including for example, plasmids, cosmids, chromosomes, phage, virus, and the like, which is capable of replication when associated with proper control elements and which can transfer polynucleotide sequences between cells. Vectors contain a nucleotide sequence that permits the vector to replicate in a selected host cell.
[0208] The term "operably linked," as used herein, refers to when transcription under the control of the "operably linked" promoter produces a functional messenger RNA, translation of which results in the production of the polypeptide encoded by the DNA operably linked to the promoter.
5.5 Fusion Proteins
[0209] Also provided herein are fusion proteins. In one embodiment, the polypeptides as provided herein, or fragments thereof, are recombinantly fused or chemically conjugated (e.g., covalent and non-covalent conjugations) to heterologous polypeptides (i.e., an unrelated polypeptide or portion thereof, preferably at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90 or at least 100 amino acids of the polypeptide) to generate fusion proteins. The fusion can be direct, or may occur through linker sequences.
[0210] In one embodiment, the fusion protein comprises a polypeptide fused to a heterologous signal sequence at its N-terminus. For example, the signal sequence naturally found in the polypeptide can be replaced by a signal sequence that is derived from a heterologous origin. Various signal sequences are commercially available.
[0211] In another embodiment, a polypeptide can be fused to tag sequences, e.g., a hexa-histidine peptide, among others, many of which are commercially available. As described in Gentz et al., 1989, Proc. Natl. Acad. Sci. USA, 86:821-824, for instance, hexa-histidine provides for convenient purification of the fusion protein. Other examples of peptide tag include the hemagglutinin "HA" tag, which corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., 1984, Cell, 37:767), and the "flag" tag (Knappik et al., 1994, Biotechniques, 17(4):754-761). These tags are useful for purification of recombinantly produced polypeptides.
[0212] Fusion proteins can be produced by standard recombinant DNA techniques or by protein synthetic techniques, e.g., by use of a DNA synthesizer. For example, a nucleic acid molecule encoding a fusion protein can be synthesized by conventional techniques including, for example, automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers, which give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, e.g., Current Protocols in Molecular Biology, Ausubel et al., eds., John Wiley & Sons, 1992).
[0213] The nucleotide sequence encoding a fusion protein can be inserted into an appropriate expression vector, i.e., a vector that contains the necessary elements for the transcription and translation of the inserted protein-coding sequence. In a specific embodiment, the expression of a fusion protein is regulated by an inducible promoter.
[0214] In another embodiment, the present invention provides methods for detecting the presence, activity or expression of a polypeptide of the invention or similar polypeptide in a biological material, such as cells, or culture media. The increased or decreased activity or expression of the polypeptide in a sample relative to a control sample can be determined by contacting the biological material with an agent that can detect directly or indirectly the presence, activity or expression of the polypeptide. In a particular embodiment, such an agent is an antibody or a fragment thereof which immunospecifically binds to one of the disclosed polypeptides.
[0215] In a still another embodiment, a fusion protein comprising a bioactive molecule and one or more domains of a disclosed polypeptide or fragment thereof is provided. In particular, fusion proteins comprising a bioactive molecule recombinantly fused or chemically conjugated (including both covalent and non-covalent conjugations) to one or more domains of a disclosed polypeptide or fragments thereof.
5.6 Preparation of Transgenic Plants
[0216] Genetic engineering of plants can be achieved in several ways. The most common method is Agrobacterium-mediated transformation. In this method, A. tumefaciens, which naturally infects plants by inserting tumor-causing genes into a plant's genome, is genetically altered. Selected genes can be engineered into the T-DNA of the bacterial Ti (tumor-inducing) plasmid of A. tumefaciens in laboratory conditions so that they become integrated into the plant chromosomes when the T-DNA is transferred to the plant by the bacteria's own internal transfer mechanisms.
[0217] The only essential parts of the T-DNA are its two small (25 base pair) border repeats, at least one of which is needed for plant transformation. The bacterial genes encoding for plant hormones that promote tumor growth are excised from the T-DNA and replaced with a sequence of DNA that typically contains: a selectable marker (e.g. an antibiotic-resistance gene; usually kanamycin resistance), a restriction site--a site with a specific sequence of nucleotides where a restriction enzyme will cut the DNA, and the desired genes to be incorporated into the plant (B. Tinland, 1996. The integration of T-DNA into plant genomes. Trends in Plant Science 1,178-184; D. Grierson (ed.) 1991. Plant Genetic Engineering. Blackie, Glasgow).
[0218] Agrobacterium can be added to plant protoplasts (plant cells with cell walls removed) in culture; the plant protoplasts then regenerate cell walls at which point non-transformed plants are killed with antibiotics for which the transformed plants have been given resistance genes. Plantlets are then regenerated from the surviving transformed cells using standard plant tissue culture techniques.
[0219] In an alternative technique, sterile disks or fragments of vegetative portions of plants are placed in liquid culture medium with Agrobacterium, and then hormones are used to induce rooting, thereby regenerating plantlets grown on selection media. Another technique for delivering genes is possible for some plants such as Arabidopsis, where the Agrobacterium or even "naked" DNA can be infused through the seed coat to cause transformation (Clough S J and Bent A F, 1998. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735-43).
[0220] The biolistic method for genetic engineering of plants was developed more recently and is becoming more widely employed. In this method, very small particles (microprojectiles) of tungsten or gold coated with biologically active DNA are propelled at high-velocities into plant cells using an electrostatic pulse, air pressure, or gunpowder percussion. As the particles pass through the cell, the DNA dissolves and can then integrate into the genome of that cell and its progeny. This method can produce stable transformants (Christou, P., et al., 1988. Stable transformation of soybean callus by DNA-coated gold particles, Plant Physiology 87:671-674). The method can be practiced on whole plants and is particularly effective on meristematic tissue. It is also capable of delivering DNA either to the nucleus or into mitochondria (Johnston, S. A., et al., 1988. Mitochondrial transformation in yeast by bombardment with microprojectiles (Science 240,1538-41) and chloroplasts (Svab, Z., et al., 1990, Stable transformation of plastids in higher plants, Proc Natl Acad Sci. USA 87, 8526-8530).
[0221] The electroporation method of plant genetic engineering has met with less success. In this technique, protoplasts in culture take up pure DNA when treated with certain membrane-active agents or with electroporation--a rapid pulse of high-voltage direct current. Once the DNA enters the protoplast, it can be integrated into the cells genome. Standard tissue culture techniques are then used to regenerate transgenic plants.
[0222] The microinjection method of plant genetic engineering is perhaps the most difficult. In this method, DNA is microinjected into target plant cells using very thin glass needles in a method similar to that used with animals. The technique is laborious, ineffective, and impractical for generating large numbers of transgenic plants.
[0223] It is within the ability of a skilled artisan to select known methods for producing genetically engineering plants, taking into account various factors such as the targeted plant species and which methods have been proven effective therein.
5.7 Preparation of Antibodies
[0224] In one aspect, provided herein are antibodies against the kinase and phosphatase. Antibodies which specifically recognize one of the described phosphatase polypeptides or fragments thereof can be used for detecting, screening, and isolating the polypeptide that is provided herein or fragments thereof, or similar sequences that encode similar enzymes from other organisms. For example, an antibody which immunospecifically binds a protein or protein fragments thereof can be used for various in vitro detection assays, including enzyme-linked immunosorbent assays (ELISA), radioimmunoassays, Western blot, etc., for the detection of the polypeptide that is provided herein or fragments, derivatives, homologues, or variants thereof, or similar molecules having the similar enzymatic activities as the phosphatase and/or kinase polypeptides.
[0225] Embodiments further provide antibodies that immunospecifically bind a polypeptide that is provided herein. Such antibodies include, but are not limited to, antibodies from various animals, humanized, chimeric, polyclonal, monoclonal, bi-specific, multi-specific, single chain antibodies, Fab fragments, F(ab')2 fragments, disulfide-linked Fvs, fragments containing a VL or VH domain or a complementary determining region (CDR), wherein the antibody or antibody fragment immunospecifically binds to a polypeptide that is provided herein.
[0226] Antibodies specific for the described phosphatase polypeptides can be generated by any suitable method known in the art. Once an antibody molecule has been produced, it may then be purified by any method known in the art for purification of an immunoglobulin molecule, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A or Protein G purification, and sizing column chromatography), centrifugation, differential solubility, or by any other standard techniques for the purification of proteins. Further, the antibodies or fragments thereof may be fused to heterologous polypeptide sequences described herein or otherwise known in the art to facilitate purification.
[0227] Antibodies fused or conjugated to heterologous polypeptides may be used in in vitro immunoassays and in purification methods (e.g., affinity chromatography) well known in the art. See e.g., PCT publication Number WO 93/21232; EP 439,095; Naramura et al., 1994, Immunol. Lett. 39:91-99; U.S. Pat. No. 5,474,981; Gillies et al., 1992, PNAS 89:1428-1432; and Fell et al., 1991, J. Immunol. 146:2446-2452, which are incorporated herein by reference in their entireties.
[0228] Antibodies may also be attached to solid supports, which are particularly useful for immunoassays or purification of the described polypeptides or fragments, derivatives, homologues, or variants thereof, or similar molecules having the similar enzymatic activities as the polypeptide of the invention. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, and polystyrene.
5.8 Detection Assays
[0229] An exemplary method for detecting the presence or absence of an over-expressed phosphatase/kinase polypeptide or an inserted phosphatase/kinase-encoding nucleic acid in a biological sample involves obtaining a biological sample from various sources and contacting the sample with a compound or an agent capable of detecting a polypeptide or nucleic acid (e.g., mRNA, genomic DNA) such that the presence of a heterologous polypeptide or nucleic acid is detected in the sample.
[0230] An exemplary agent for detecting mRNA or genomic DNA encoding an inserted phosphatase polypeptide is a labeled nucleic acid probe capable of hybridizing to mRNA or genomic DNA encoding any of the described phosphatase and kinase polypeptides. The nucleic acid probe can be, for example, a full-length cDNA, such as the nucleic acid of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 93, 95, 97, 99, 101 or a portion thereof, such as an oligonucleotide of at least one of at least about 15, at least about 20, at least about 25, at least about 30, at least about 50, at least about 100, at least about 250, at least about 500, or more nucleotides in length and sufficient to specifically hybridize under stringent conditions to a mRNA or genomic DNA encoding a polypeptide of the invention.
[0231] An exemplary agent for detecting an over-expressed phosphatase/kinase polypeptide is an antibody capable of binding to a phosphatase/kinase polypeptide product of an inserted gene, preferably an antibody with a detectable label. Antibodies can be polyclonal and monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab')2) can be used.
[0232] The term "labeled", with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently labeled streptavidin.
[0233] The detection method can be used to detect mRNA, protein, or genomic DNA in a sample in vitro as well as in vivo. For example, in vitro techniques for detection of mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of a heterologous polypeptide include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations and immunofluorescence. In vitro techniques for detection of genomic DNA include Southern hybridizations. Furthermore, in vivo techniques for detection of a heterologous polypeptide include introducing into a subject organism a labeled antibody directed against the polypeptide. For example, the antibody can be labeled with a radioactive marker whose presence and location in the subject organism can be detected by standard imaging techniques, including autoradiography.
[0234] In a specific embodiment, the methods further involve: 1) obtaining a control sample from a control subject, 2) contacting the control sample with a compound or agent capable of detecting an over-expressed polypeptide product, or the mRNA transcription product, or genomic DNA encoding an inserted phospatase gene, such that the presence of the polypeptide or mRNA or genomic DNA encoding the phosphatase polypeptide is detected in the sample, and 3) comparing the level of the phosphatase/kinase polypeptide or mRNA or genomic DNA encoding the polypeptide in a control sample with the level of the polypeptide or mRNA or genomic DNA encoding endogenous phosphatase polypeptides in the test sample.
5.9 Applications of Transgenic Plants
[0235] The transgenic plants generated can have many useful applications, including in food, feed, biomass, biofuels (starch, cellulose, seed lipids) and wood pulp industry. The enhanced growth rate of the transgenic plants can provide additional carbon dioxide fixation per hectare of land per year, and, thus is useful for generating carbon credits.
6.0 EXAMPLES
[0236] Following are examples that illustrate embodiments for practicing the invention. These examples should not be construed as limiting. Unless otherwise noted, all percentages are by weight, all solvent mixture proportions are by volume, all temperatures are in Centigrade, and all pressure is at or near atmospheric pressure.
6.1 Screening of Grow-Promoting NG Genes
[0237] Two independent AtPAP2 overexpression lines (OE7 and OE21, homozygous T3 plants), an AtPAP2 T-DNA mutant line that cannot express the full length AtPAP2, and the wild-type Arabidopsis (Col-0) were employed for microarray analysis. The AtPAP2 overexpression lines (OE7 and OE21, homozygous T3 plants), the AtPAP2 T-DNA mutant line, and the wild-type Arabidopsis (Col-0) line have been disclosed by the present inventor in U.S. patent application Ser. No. 12/640,674 (U.S. Patent Application Publication No. 2010/0159065), which is hereby incorporated by reference in its entirety.
[0238] Briefly, seeds were germinated on MS medium supplemented with 2% (w/v) sucrose, grew in a growth room under 12 hour-light/12 hour-dark cycle at 22° C. for 10 days, and were then transferred to soil and grew in a growth chamber under a 16-hour light (22° C.) and 8-hour dark (18° C.) cycle. Shoots of 20-day-old Arabidopsis (WT, T-DNA, OE7 and OE21) prior to bolting were collected in the middle of day (4 plants/line/tube, 3 biological triplicates/line, 3 tubes/line) and ground in liquid nitrogen. RNA extraction was performed with on-column DNase digestion according to the manufacturer's instruction (RNeasy Plant Mini Kit, Cat. No. 74904, Qiagen). Total RNA was dissolved in DEPC water and quantified by the Bioanalyzer 2100 (Agilent Technologies, Boblingen, Germany). Double strand DNA synthesis and Cy 3 labeling from three biological replicates were performed by NimbleGen Systems, Inc. (Madison, Wis.). Statistical analyses of normalized microarray data (RMA algorithm, quantile normalization) and drawing of scatter plots, heatmaps were performed using ArrayStar 3.0 (DNASTAR, Madison, Wis.). Identification of GO and classification were carried out using software available from TAIR database and KEGG pathway database. In all three replications, genes were considered to be significantly regulated if their fold change values were positively or negatively beyond 1.3 (p<0.05).
[0239] 20-day-old plants did not show any differences in appearance so that any differences in gene expression between the lines were not due to difference in developmental stage or additional tissues (e.g. inflorescence). The transcripts levels of 30360 genes in shoots were determined using the Arabidopsis Genome NimbleGen chips. The average hybridization signals detected in each line were normalized from the log2 average signal and compared with the signal strengths in the wild-type Arabidopsis.
[0240] An overview of the expression data of OE7, OE21 and T-DNA plants versus wild-type control is presented as a heat map (FIG. 1) and scatter plots (FIG. 2) that show a linear bias in the graphs. Gene expression patterns in transgenic shoots are different comparative to their wild-type controls.
[0241] The data show that AtPAP2 overexpression altered expression levels of other genes, nearly half of which have not been characterized yet. AtPAP2 overexpression lines exhibit more dramatic changes in gene expression than the AtPAP2 T-DNA line.
[0242] Differentially expressed genes are identified using P-value<0.05 and fold change>1.3 as the cutoff, and the results show that the expression of about 6312, 7831, and 672 genes in the shoots of OE7, OE21 and T-DNA lines are significantly altered. An overall view of the altered genes in the heat map (FIG. 1) revealed that most genes were down-regulated in the fold change>=2.0. In addition, the fold change in expression levels is smaller in up-regulated genes than in down-regulated genes.
[0243] Based on the microarray data, 33 putative phosphatase and kinase genes were selected, and were introduced into Arabidopsis to produce overexpression lines. The results show that the overexpression of NG6, NG21, NG24, NG28 and NG32 in Arabidopsis promotes the growth of Arabidopsis and increases seed yield (see Example 2). The expression level of the five growth-promoting NG genes in the AtPAP2 OE lines and T-DNA lines are shown in Table 1.
TABLE-US-00001 TABLE 1 Microarray data of the 5 growth-promoting genes in AtPAP2 overexpression lines (OE7, OE21), T-DNA line and wild type (WT) Arabidopsis. WT- T-DNA OE7 OE21 OE7/WT OE21/WT T-DNA/WT Gene NG No. AGI code Mean Mean Mean Mean Fold Fold Fold Description NG 6 AT1G05000 637 533 976 1131 1.52* 1.78** 0.83 Protein phosphatase NG21 AT1G13350 2406 2151 3543 3441 1.47* 1.43* 0.89 Protein kinase NG24 AT1G28390 778 710 1853 1915 2.37** 2.42** 0.91 Protein kinase NG28 AT3G24660 2514 1839 3313 4422 1.32** 1.74** 0.73 Protein kinase NG32 AT5G03320 1325 1063 1884 2053 1.43* 1.56** 0.80 Protein kinase
6.2 Production of NG Overexpression Lines in Arabidopsis
[0244] To create transgenic NG gene overexpressing lines, the full length coding region of each NG gene's cDNA was amplified by PCR using the following primers (Table 2). The PCR products were inserted into the pCXSN vector with classical TA cloning method (FIG. 3).
TABLE-US-00002 TABLE 2 Primers used for to amplify the full CDS of the aimed NG genes Gene name Sequence(5'-3') NG6 Forward Primer 5'-TCGAGCTAGCATGAAGCTTGTGGAGAAGAC-3' (SEQ ID NO: 51) Reverse Primer 5'-CGACGAGCTCTTACCTGATGGAACAAGAG-3 (SEQ ID NO: 52) NG21 Forward Primer 5'-ATGGTGAGTGACAAGCATGTAG-3' (SEQ ID NO: 53) Reverse Primer 5'-TCACTTGCCCGTGATGAATG-3' (SEQ ID NO: 54) NG24 Forward Primer 5'-ATGGGTTATCTCTCTTGCAAC-3' (SEQ ID NO: 55) Reverse Primer 5'-TCAGTATCTCTTCCGCGACG-3' (SEQ ID NO: 56) NG28 Forward Primer 5'-ATGGGCATGGAAGCTTTGAG-3' (SEQ ID NO: 57) Reverse Primer 5'-TCAAAATGGAGTTTCGGCGT-3' (SEQ ID NO: 58) NG32 Forward Primer 5'-ATGAAATGCTTCTTATTCCC-3' (SEQ ID NO: 59) Reverse Primer 5'-TCAACAAGCTCTCACATTCT-3' (SEQ ID NO: 60)
[0245] The vector was introduced into Agrobacterium tumefaciens strain GV3101 and then transformed by the floral dip method (Clough and Bent, 1998) into wild-type Col-0 to generate NG-overexpressing lines. Through two generations of selection on MS agar plate with 30 mg/1 hygromycin, homologous NG transgenic lines were obtained. The resistant plants were transferred to soil to grow to maturity, and their transgenic status was confirmed by qRT-PCR analysis.
6.3 Confirmation of Overexpression of NG Genes in Transgenic Plants
[0246] The transcription levels of the NG genes in the hygromycin resistant, homologous T3 overexpression lines were confirmed by quantitative Real Time-PCR. Total RNA was extracted from 10-day-old seedlings grown on Murashige and Skoog (MS) with 3% (w/v) sucrose using the TRIzol RNA isolation method with DNase I treatment. cDNAs were generated using Superscript III reverse transcriptase (Invitrogen, Carlsbad, Calif., USA) using an oligol 5 dT primer. Two gene-specific primers were used to amplify the 80-150 bp coding region of each NG gene. The ACTIN primers were used for control experiment. As shown in FIG. 4, the transcript levels of each overexpression line were consistently higher than their respective expression levels in the wild-type.
TABLE-US-00003 TABLE 3 Primers used in the quantitative RT-PCR NG6 Forward Primer 5'-TGTGCCCGGAGCCCTACC-3' (SEQ ID NO: 61) Reverse Primer 5'-CTTTCAGTGCCATGCGGATTTT-3 (SEQ ID NO: 62) NG21 Forward Primer 5'-GGCACAAGTCCCGTCATCACC-3' (SEQ ID NO: 63) Reverse Primer 5'-TCCCCAATCCCTTCTTTTCCTA-3' (SEQ ID NO: 64) NG24 Forward Primer 5'-GCCGCCGTCAAGAGAACAAC-3' (SEQ ID NO: 65) Reverse Primer 5'-CTCCGGTGGTCAACGCAGTAA-3' (SEQ ID NO: 66) NG28 Forward Primer 5'-TGTTGTTGTGGCCTCGTTGTTA-3' (SEQ ID NO: 67) Reverse Primer 5'-CTTTCCTTCACCGCCTTCTTTC-3' (SEQ ID NO: 68) NG32 Forward Primer 5'-AAGCTTTCGGATTTCGGTTTG-3' (SEQ ID NO: 69) Reverse Primer 5'-TGGCCTTCTTCCTGTAATGAGC-3' (SEQ ID NO: 70) ACTIN Forward Primer 5'-CCCGCTATGTATGTCGC-3' (SEQ ID NO: 71) Reverse Primer 5'-AAGGTCAAGACGGAGGAT-3' (SEQ ID NO: 72)
6.4 Growth Phenotypes of NG Gene Over-Expression Lines
[0247] Arabidopsis seeds were soaked in water at 4° C. for 3 days. The seeds were surface sterilized and sown on MS medium supplemented with 3% (w/v) sucrose for 10 days. Seedlings with 2 rosette leaves of the same size were transferred to soil under Long Day condition (16 h light at 22° C./8 h dark at 18° C.) in a plant growth chamber. Bolting time was measured when the primary inflorescence reached 1 cm above the rosette leaves. (Liu et al., 2008; Wu et al., 2008).
[0248] The inflorescences of NG gene OE lines emerged earlier (4-5 days) than the WT at Long Day conditions (Table 4, FIG. 5 and FIG. 6). This phenotype observation was repeated at least 3 times and the results of two of the experiments are shown here.
TABLE-US-00004 TABLE 4 WT NG6 NG21 NG24 NG28 NG32 A. Earlier bolting time of NG OE lines (Trial 1) Average bolting 24.4 21.2 20.1 21.4 20.8 19.8 time (Day) SD 1.4 1.0 1.4 0.9 1.0 1.3 N 12 12 12 9 9 9 B. Earlier bolting time of NG OE lines (Trial 2) Average bolting 24.3 19.2 19 19 18.3 19 time (Day) SD 0.8 0.8 1.1 1 1.0 0.9 N 12 6 6 9 6 6
[0249] At maturity (Long Day), the number of inflorescence and the total weight of seeds harvested from each line were recorded. The results of two separate experimental trials are shown in Tables 5A and B. The results show that the overexpression of each of the five NG genes (NG6, NG21, NG24, NG28, and NG32) resulted in increased number of inflorescences and seed yield. Compared to that of the wild-type, the seed yield of each NG over-expression line increased 30-50% (Table 5).
TABLE-US-00005 TABLE 5A OE lines produced more seeds (Trial 1). Lines Weight of seeds (mg)/plant SD WT(Col-0) 80.4 4.9 NG6 113.6 12.2 NG21 127.8 26.9 NG24 99.6 17.3 NG28 130.6 26.7 NG32 135.9 23.5
The plants were grown in small black trays (N=6-9).
TABLE-US-00006 TABLE 5B OE lines produced more seeds (Trial 2). Lines Weight of seeds (mg)/plant SD WT(Col-0) 142.0 14.6 NG6 190.3 15.7 NG21 180.4 26.3 NG24 203.8 20.0 NG28 186.0 39.5 NG32 241.8 23.8
The plants were grown in large white cups (N=6-9).
[0250] The results show that, when compared to the wild-type, Arabidopsis plants transformed with NG6, NG21, NG24, NG28 and/or NG32 have the following advantageous phenotypes: (1) faster growth rate; (2) higher seed yield.
6.5 Sequence Alignment and Phylogenetic Analysis
[0251] All the CDS of 5 NG genes in the Arabidopsis Col-0 ecotype were obtained from the TAIR website. Sequence alignment of each NG gene was retrieved by tblastn program from Plant GDB database and NCBI database using the amino acid sequence of each Arabidopsis NG gene as the bait sequences. Partial sequences recovered were aligned and compared to produce a full length coding sequence if feasible. Sequence alignment and phylogenetic tree were conducted using MEGA4 (Kumar et al., 2004) and ClustalW program. Amino acid sequence comparisons were performed using CLC Sequence Viewer 5.1.1.
[0252] Those skilled in the art will recognize, or be able to ascertain many equivalents to the specific embodiments of the invention described herein using no more than routine experimentation. Such equivalents are intended to be encompassed by the following claims.
[0253] All publication, patents and patent applications mentioned in this specification are incorporated herein by reference in their entireties into the specification to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety.
[0254] Citation or discussion of a reference herein shall not be construed as an admission that such is prior art to the present invention.
REFERENCES
[0255] Clough, S. J. and Bent, A. F. (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J, 16, 735-743.
[0256] Klabunde, T., Strater, N., Frohlich, R., Witzel, H. and Krebs, B. (1996) Mechanism of Fe(III)-Zn(II) purple acid phosphatase based on crystal structures. J. mol. biol., 259, 737-748.
[0257] Klabunde, T. and Krebs, B. (1997) The dimetal center in purple acid phosphatases. Metal Sites in Proteins and Models, 89, 177-198.
[0258] Li, D., Zhu, H., Liu, K., Liu, X., Leggewie, G., Udvardi, M. and Wang, D. (2002) Purple acid phosphatases of Arabidopsis thaliana. Comparative analysis and differential regulation by phosphate deprivation. J. Biol. Chem., 277, 27772-27781.
[0259] Schenk, G., Ge, Y., Carrington, L. E., Wynne, C. J., Searle, I. R., Carroll, B. J., Hamilton, S. and de-Jersey, J. (1999) Binuclear metal centers in plant purple acid phosphatases: Fe--Mn in sweet potato and Fe-Zn in soybean. Arch. Biochem Biophys, 370, 183-189.
[0260] United States Patent Application Publication No. 2010/0159065
Sequence CWU
1
1
1021648DNAArabidopsis thaliana 1atgaagcttg tggagaagac gactactaca
gagcaggaca atggagaaga tttctgccgc 60accatcatcg aggtttccga ggttaacaga
aacgtgtttc aggctccggg cggtgaagct 120gatccctttc gagttgtctc cggcgaagaa
cttcacctaa ttccgccgct caacttctcc 180atggtcgata acggtatatt ccggtctgga
ttccctgatt cagctaactt ctcctttctc 240cagactctcg gtctccgctc aatcatatac
ttgtgcccgg agccctaccc agagagcaat 300ctccagttcc ttaaatccaa tggaatcagg
cttttccagt ttggtattga aggcaacaag 360gagccatttg tgaatattcc agaccataaa
atccgcatgg cactgaaagt gcttctagat 420gagaaaaacc atcctgttct gattcattgt
aagcgaggca agcatcggac cggttgtctt 480gttggttgct tgaggaagct ccagaaatgg
tgtttgacat cgatattcga cgagtaccaa 540cgatttgcag cagcgaaagc tagagtttca
gatcaaagat tcatggagat attcgatgtt 600tccagcttca gtcatattcc aatgtcattc
tcttgttcca tcaggtaa 6482215PRTArabidopsis thaliana 2Met
Lys Leu Val Glu Lys Thr Thr Thr Thr Glu Gln Asp Asn Gly Glu 1
5 10 15 Asp Phe Cys Arg Thr Ile
Ile Glu Val Ser Glu Val Asn Arg Asn Val 20
25 30 Phe Gln Ala Pro Gly Gly Glu Ala Asp Pro
Phe Arg Val Val Ser Gly 35 40
45 Glu Glu Leu His Leu Ile Pro Pro Leu Asn Phe Ser Met Val
Asp Asn 50 55 60
Gly Ile Phe Arg Ser Gly Phe Pro Asp Ser Ala Asn Phe Ser Phe Leu 65
70 75 80 Gln Thr Leu Gly Leu
Arg Ser Ile Ile Tyr Leu Cys Pro Glu Pro Tyr 85
90 95 Pro Glu Ser Asn Leu Gln Phe Leu Lys Ser
Asn Gly Ile Arg Leu Phe 100 105
110 Gln Phe Gly Ile Glu Gly Asn Lys Glu Pro Phe Val Asn Ile Pro
Asp 115 120 125 His
Lys Ile Arg Met Ala Leu Lys Val Leu Leu Asp Glu Lys Asn His 130
135 140 Pro Val Leu Ile His Cys
Lys Arg Gly Lys His Arg Thr Gly Cys Leu 145 150
155 160 Val Gly Cys Leu Arg Lys Leu Gln Lys Trp Cys
Leu Thr Ser Ile Phe 165 170
175 Asp Glu Tyr Gln Arg Phe Ala Ala Ala Lys Ala Arg Val Ser Asp Gln
180 185 190 Arg Phe
Met Glu Ile Phe Asp Val Ser Ser Phe Ser His Ile Pro Met 195
200 205 Ser Phe Ser Cys Ser Ile Arg
210 215 3651DNAZea mays 3atgaagctgg aggtcatgcc
caagcccaaa cagcgggtgc tggaggcgca gcagagggag 60gaggccatgg agatgagcgg
cctggacctg tggaagcacg agaagccgcc aaggatctgc 120cccttgcccc cgtcgctccc
gccgccgccg ccagcgttcg acgaggcggc gctcgtgccg 180ccgctcaact tcgccgtggt
cgacgacggc atcttccgct ccggattccc agggaccgcc 240aacttccggt tcctcaagtc
cctcaacctc cgctccatcg tgtacctgtg cccggagccg 300tacccgggga cgaacacgga
gttcctagaa aagaatggga tcaggctcca ccagttcgga 360atcgaggggc gcaaggaacc
atttgtcaac atacccgacg acaaaataag ggaggcgctc 420aaagttgtct tagacccaag
aaaccaacct ctgcttatcc attgcaagag aggcaagcac 480cgaactggct gtgtggtcgg
atgcttgagg aagctgcagg aatggtgctt gtcttcagtc 540ttggacgagt accatcgctt
tgccgctgcg aaagcgagga tcactgacca gaggttcatg 600gagctgttcg acgtttcaag
cttgaagcac ctgacaccct cacactgtta a 6514216PRTZea mays 4Met
Lys Leu Glu Val Met Pro Lys Pro Lys Gln Arg Val Leu Glu Ala 1
5 10 15 Gln Gln Arg Glu Glu Ala
Met Glu Met Ser Gly Leu Asp Leu Trp Lys 20
25 30 His Glu Lys Pro Pro Arg Ile Cys Pro Leu
Pro Pro Ser Leu Pro Pro 35 40
45 Pro Pro Pro Ala Phe Asp Glu Ala Ala Leu Val Pro Pro Leu
Asn Phe 50 55 60
Ala Val Val Asp Asp Gly Ile Phe Arg Ser Gly Phe Pro Gly Thr Ala 65
70 75 80 Asn Phe Arg Phe Leu
Lys Ser Leu Asn Leu Arg Ser Ile Val Tyr Leu 85
90 95 Cys Pro Glu Pro Tyr Pro Gly Thr Asn Thr
Glu Phe Leu Glu Lys Asn 100 105
110 Gly Ile Arg Leu His Gln Phe Gly Ile Glu Gly Arg Lys Glu Pro
Phe 115 120 125 Val
Asn Ile Pro Asp Asp Lys Ile Arg Glu Ala Leu Lys Val Val Leu 130
135 140 Asp Pro Arg Asn Gln Pro
Leu Leu Ile His Cys Lys Arg Gly Lys His 145 150
155 160 Arg Thr Gly Cys Val Val Gly Cys Leu Arg Lys
Leu Gln Glu Trp Cys 165 170
175 Leu Ser Ser Val Leu Asp Glu Tyr His Arg Phe Ala Ala Ala Lys Ala
180 185 190 Arg Ile
Thr Asp Gln Arg Phe Met Glu Leu Phe Asp Val Ser Ser Leu 195
200 205 Lys His Leu Thr Pro Ser His
Cys 210 215 5627DNAglycine max 5atgcaagtgg
cggcagaact ccaacgcgcc catcaccacc accaaaggca caaacaagac 60actcccatgt
gccgccaaat ccagctcact atctccgatc acaccaccgc cggagacgac 120gacggcgagg
atctcttcat tccgcccctc aacttcgcca tggttgataa tggcattttc 180cgctccggct
tccccgaacc cgccaacttc tccttcctcc aaaccctcgg cctccgttcc 240atcatatatc
tgtgtcctga gccgtatccg gaggccaata tggagttcct caagtcaaat 300gggatcaagc
tttttcagtt tgggattgag ggtcataagg agccttttgt gaacatccca 360gaggacacaa
tccgtgaagc actaaaagtt gttcttgatg tcaggaacca cccagttata 420attcactgta
agcgtggaaa gcaccgaacg ggttgcttag taggatgcta tagaaaattg 480caaaaatggt
gcttgtcatc tgtctttgat gaataccaac gctttgcagc tgccaaagca 540agagtttcag
atcagaggtt tgtagagttg tttgatattt ccagcctgaa acattttcct 600ataccctttt
catgtttgaa gaggtga
6276208PRTglycine max 6Met Gln Val Ala Ala Glu Leu Gln Arg Ala His His
His His Gln Arg 1 5 10
15 His Lys Gln Asp Thr Pro Met Cys Arg Gln Ile Gln Leu Thr Ile Ser
20 25 30 Asp His Thr
Thr Ala Gly Asp Asp Asp Gly Glu Asp Leu Phe Ile Pro 35
40 45 Pro Leu Asn Phe Ala Met Val Asp
Asn Gly Ile Phe Arg Ser Gly Phe 50 55
60 Pro Glu Pro Ala Asn Phe Ser Phe Leu Gln Thr Leu Gly
Leu Arg Ser 65 70 75
80 Ile Ile Tyr Leu Cys Pro Glu Pro Tyr Pro Glu Ala Asn Met Glu Phe
85 90 95 Leu Lys Ser Asn
Gly Ile Lys Leu Phe Gln Phe Gly Ile Glu Gly His 100
105 110 Lys Glu Pro Phe Val Asn Ile Pro Glu
Asp Thr Ile Arg Glu Ala Leu 115 120
125 Lys Val Val Leu Asp Val Arg Asn His Pro Val Ile Ile His
Cys Lys 130 135 140
Arg Gly Lys His Arg Thr Gly Cys Leu Val Gly Cys Tyr Arg Lys Leu 145
150 155 160 Gln Lys Trp Cys Leu
Ser Ser Val Phe Asp Glu Tyr Gln Arg Phe Ala 165
170 175 Ala Ala Lys Ala Arg Val Ser Asp Gln Arg
Phe Val Glu Leu Phe Asp 180 185
190 Ile Ser Ser Leu Lys His Phe Pro Ile Pro Phe Ser Cys Leu Lys
Arg 195 200 205
7678DNAOryza sativa 7atgaagctgg aggtgatgcc gaagcagagg gccatggagg
ccgagcagag ggaggaggcc 60atggagatga gcgggctcga gctgtggaag cacgagaagc
ccgcgtccat ggtggtgttc 120ctcccgccgc cgccgccgcc gccgcttgtg ccggcggcgg
cggccgcggc cgcggcggcg 180tgtggtgagg aggcgacgct ggtgccaccg ctcaacttcg
cgatggtcga cgacggcatc 240ttccgctccg gcttccccgc ggccgccaac ttccggttcc
tcaagtcgct caacctccgc 300tccatcgtgt acctgtgccc ggagccgtac ccggagacga
acgcggagtt cctcgccaag 360aacgggatca agctccacca gttcggaatc gaggggcgca
aggaaccatt cgtcaacatc 420cctgacgaca aaattcgaga ggcgctcaaa gttgtcctag
acgtaaaaaa ccaacctctg 480cttattcact gcaagagagg caagcaccgc accggctgcg
tcgtggggtg cttgaggaag 540cttcagaaat ggtgcttgtc ttcagtgttc gacgagtacc
agcgcttcgc cgctgcgaag 600gcgaggagca ccgatcagag attcatggag ctgttcgaca
tctcaagctt gaagcacctg 660acagcttcac attgttaa
6788225PRToryza sativa 8Met Lys Leu Glu Val Met
Pro Lys Gln Arg Ala Met Glu Ala Glu Gln 1 5
10 15 Arg Glu Glu Ala Met Glu Met Ser Gly Leu Glu
Leu Trp Lys His Glu 20 25
30 Lys Pro Ala Ser Met Val Val Phe Leu Pro Pro Pro Pro Pro Pro
Pro 35 40 45 Leu
Val Pro Ala Ala Ala Ala Ala Ala Ala Ala Ala Cys Gly Glu Glu 50
55 60 Ala Thr Leu Val Pro Pro
Leu Asn Phe Ala Met Val Asp Asp Gly Ile 65 70
75 80 Phe Arg Ser Gly Phe Pro Ala Ala Ala Asn Phe
Arg Phe Leu Lys Ser 85 90
95 Leu Asn Leu Arg Ser Ile Val Tyr Leu Cys Pro Glu Pro Tyr Pro Glu
100 105 110 Thr Asn
Ala Glu Phe Leu Ala Lys Asn Gly Ile Lys Leu His Gln Phe 115
120 125 Gly Ile Glu Gly Arg Lys Glu
Pro Phe Val Asn Ile Pro Asp Asp Lys 130 135
140 Ile Arg Glu Ala Leu Lys Val Val Leu Asp Val Lys
Asn Gln Pro Leu 145 150 155
160 Leu Ile His Cys Lys Arg Gly Lys His Arg Thr Gly Cys Val Val Gly
165 170 175 Cys Leu Arg
Lys Leu Gln Lys Trp Cys Leu Ser Ser Val Phe Asp Glu 180
185 190 Tyr Gln Arg Phe Ala Ala Ala Lys
Ala Arg Ser Thr Asp Gln Arg Phe 195 200
205 Met Glu Leu Phe Asp Ile Ser Ser Leu Lys His Leu Thr
Ala Ser His 210 215 220
Cys 225 9537DNAGossypium hirsutum 9atgtgcagaa ccatagaaga agatgccctc
gccgttgacc accacgtcga catgtcgtcg 60tcgacttcag atgacctcaa cttgattcct
cctttgaact ttgctatagt tgacaatggc 120atcttcaggt ctggtttccc tgattctgcc
aacttctctt ttcttcaaac gcttaagctc 180acctccatca tatatctgtg tcctgaacca
tacccagaag ccaacactga gtttttaaag 240tccgatggaa tcaagctttt tcagtttgga
attgaaagtt acaaggagcc atttgtaaat 300attccagagg atacgattcg tgaagcttta
aggctcgtcc tcgatgttag gaatcaccca 360gttttaattc attgtaatcg agggaagcac
cgaactggtc gtctggttgg atgcctgagg 420aagttgcaga gatggtgttt gtcatccgtg
ttcgacgagt accaaaggct tgctgccgca 480aaagctagag tttcggatca gagcggagaa
tgctcggtcc tgcataggat taattag 53710178PRTGossypium hirsutum 10Met
Cys Arg Thr Ile Glu Glu Asp Ala Leu Ala Val Asp His His Val 1
5 10 15 Asp Met Ser Ser Ser Thr
Ser Asp Asp Leu Asn Leu Ile Pro Pro Leu 20
25 30 Asn Phe Ala Ile Val Asp Asn Gly Ile Phe
Arg Ser Gly Phe Pro Asp 35 40
45 Ser Ala Asn Phe Ser Phe Leu Gln Thr Leu Lys Leu Thr Ser
Ile Ile 50 55 60
Tyr Leu Cys Pro Glu Pro Tyr Pro Glu Ala Asn Thr Glu Phe Leu Lys 65
70 75 80 Ser Asp Gly Ile Lys
Leu Phe Gln Phe Gly Ile Glu Ser Tyr Lys Glu 85
90 95 Pro Phe Val Asn Ile Pro Glu Asp Thr Ile
Arg Glu Ala Leu Arg Leu 100 105
110 Val Leu Asp Val Arg Asn His Pro Val Leu Ile His Cys Asn Arg
Gly 115 120 125 Lys
His Arg Thr Gly Arg Leu Val Gly Cys Leu Arg Lys Leu Gln Arg 130
135 140 Trp Cys Leu Ser Ser Val
Phe Asp Glu Tyr Gln Arg Leu Ala Ala Ala 145 150
155 160 Lys Ala Arg Val Ser Asp Gln Ser Gly Glu Cys
Ser Val Leu His Arg 165 170
175 Ile Asn 112286DNAArabidopsis thaliana 11atggtgagtg acaagcatgt
agaatcaaac caccgcaaac accgacggtc gttttcgccg 60tccgacgagg tctttaaatc
tccgaagcgg cacaagtccc gtcatcacca tcgcaggcat 120ggccaccgtc atcatcgtga
tgaggaagtt caatataacg atgatgagaa tgttaacggt 180ggtgatcttg atatggaaga
aggtgagata ttaggaaaag aagggattgg ggagacattg 240aagaagaaat tagagtccgt
cgacgagttt ggggatataa aatctggtca attccgggag 300aataatctgg ggagaaatca
gcggagggaa agagaatgtg agaaaagaaa agagatagag 360cctgaccgtg aaaggagaaa
agagagggga agcgttgata gagatagcag gggagacagg 420gaaaaagatt acctacggga
tagagacaac gacagaggta ggagtagaga taaagccagg 480tatagtagta gagagagggg
gagggagaat gaaagagaga gacggagtga aaaagatagg 540gataaaggac gagaattcca
gagtgataga gagaagcata aaagtcttga tgatggatat 600ggtgaagtga ggcataaaca
ttctggacac tcaagacatg atgcggaaga tgacttagag 660ttaagaagcc caacttctgt
aaatggccat gatcctaaca gtggcgatgt caaagaaact 720cggggaaatg ttgaaaggac
cagaattgat aatgatgata aaggtgacgt tgttgtttgg 780gaagttgaac aagaagatga
agagctaaat ttaatcgagg aaagcaggag gagaacgcaa 840gccataatgg agaaatataa
gaaaaagttg gagcagcaaa acggattttc ttctcatgat 900cttgagctag caaacattcc
caagcagtcc tctactgtgg cagatgttct tggaagtggt 960actctggggc ctgttacttc
tgcagttaat caagctaaag ctgggttgga tattgatgcc 1020gtagatggtg aagtcgccaa
gctttcatcg gcagttgggg aatcacctgc acagcttgta 1080atttcagact cagataggac
actagcttcc acagggcttg gggaaggcag cccaaaggat 1140aaaatatcag atgacatgtt
cactgatgat atctttgggg agtctccagc tgatagtcag 1200aaaatgggct atctgcgagg
gaaagggaat ggcattccta ttgtaaggag tggactcgac 1260gataattggg atgatgcaga
aggttattac agttatcaat taggggaact acttgatgat 1320agatatgaaa tcatggctac
tcatggaaaa ggtgtcttct ctaccgtggt gcgggcaaaa 1380gacacaaaag ctgaactagg
tgaacctgag gaagtggcta taaaaattat tcggaacaat 1440gagacaatgc ataaggccgg
ccagactgag attcagatat tgaagaagct agctggctct 1500gacccagaga ataagcgcca
ctgcgttcgt tttctttcaa cttttaagta taggaaccac 1560ctttgcttgg tgtttgagtc
tcttcatctg aatctccgtg agattgtgaa gaagtatggt 1620cgcaacattg gtattcaact
atctggtgtt agagtgtatg caacgcagtt attcatatcc 1680cttaaacatc tcaagaactg
tggggttctt cactgcgata taaagcctga caacatgctg 1740gtgaatgagg gaagaaacac
gttaaagctt tgtgactttg gtagtgcaat gtttgctggt 1800acaaacgaag ttacaccata
tcttgttagt cgcttctaca gagctccaga aataattctt 1860ggacttccct acgaccatcc
gttagatata tggtcagttg gttgctgtct gtatgagctt 1920tttagcggga aaattatgtt
ccctggctcc acaaacaatg aaatgttacg cctgcatatg 1980gaactgaaag gtgccttccc
taaaaagatg cttcgcaagg gagcatttat cgatcagcac 2040tttgataagg acttatgctt
ctatgctaca gaggaggata gtgttactag aaagacaaca 2100aagagaatga tggtaaacat
aaagccaaaa gaatttggtt cagtaattaa acaacgttat 2160aaggatgaag atagcaagtt
gttggttcat ttcagggatc ttctagacag aattttcata 2220cttgatcctc agaagagaat
tacagtgtca caggcattag ctcacccatt catcacgggc 2280aagtga
228612761PRTArabidopsis
thaliana 12Met Val Ser Asp Lys His Val Glu Ser Asn His Arg Lys His Arg
Arg 1 5 10 15 Ser
Phe Ser Pro Ser Asp Glu Val Phe Lys Ser Pro Lys Arg His Lys
20 25 30 Ser Arg His His His
Arg Arg His Gly His Arg His His Arg Asp Glu 35
40 45 Glu Val Gln Tyr Asn Asp Asp Glu Asn
Val Asn Gly Gly Asp Leu Asp 50 55
60 Met Glu Glu Gly Glu Ile Leu Gly Lys Glu Gly Ile Gly
Glu Thr Leu 65 70 75
80 Lys Lys Lys Leu Glu Ser Val Asp Glu Phe Gly Asp Ile Lys Ser Gly
85 90 95 Gln Phe Arg Glu
Asn Asn Leu Gly Arg Asn Gln Arg Arg Glu Arg Glu 100
105 110 Cys Glu Lys Arg Lys Glu Ile Glu Pro
Asp Arg Glu Arg Arg Lys Glu 115 120
125 Arg Gly Ser Val Asp Arg Asp Ser Arg Gly Asp Arg Glu Lys
Asp Tyr 130 135 140
Leu Arg Asp Arg Asp Asn Asp Arg Gly Arg Ser Arg Asp Lys Ala Arg 145
150 155 160 Tyr Ser Ser Arg Glu
Arg Gly Arg Glu Asn Glu Arg Glu Arg Arg Ser 165
170 175 Glu Lys Asp Arg Asp Lys Gly Arg Glu Phe
Gln Ser Asp Arg Glu Lys 180 185
190 His Lys Ser Leu Asp Asp Gly Tyr Gly Glu Val Arg His Lys His
Ser 195 200 205 Gly
His Ser Arg His Asp Ala Glu Asp Asp Leu Glu Leu Arg Ser Pro 210
215 220 Thr Ser Val Asn Gly His
Asp Pro Asn Ser Gly Asp Val Lys Glu Thr 225 230
235 240 Arg Gly Asn Val Glu Arg Thr Arg Ile Asp Asn
Asp Asp Lys Gly Asp 245 250
255 Val Val Val Trp Glu Val Glu Gln Glu Asp Glu Glu Leu Asn Leu Ile
260 265 270 Glu Glu
Ser Arg Arg Arg Thr Gln Ala Ile Met Glu Lys Tyr Lys Lys 275
280 285 Lys Leu Glu Gln Gln Asn Gly
Phe Ser Ser His Asp Leu Glu Leu Ala 290 295
300 Asn Ile Pro Lys Gln Ser Ser Thr Val Ala Asp Val
Leu Gly Ser Gly 305 310 315
320 Thr Leu Gly Pro Val Thr Ser Ala Val Asn Gln Ala Lys Ala Gly Leu
325 330 335 Asp Ile Asp
Ala Val Asp Gly Glu Val Ala Lys Leu Ser Ser Ala Val 340
345 350 Gly Glu Ser Pro Ala Gln Leu Val
Ile Ser Asp Ser Asp Arg Thr Leu 355 360
365 Ala Ser Thr Gly Leu Gly Glu Gly Ser Pro Lys Asp Lys
Ile Ser Asp 370 375 380
Asp Met Phe Thr Asp Asp Ile Phe Gly Glu Ser Pro Ala Asp Ser Gln 385
390 395 400 Lys Met Gly Tyr
Leu Arg Gly Lys Gly Asn Gly Ile Pro Ile Val Arg 405
410 415 Ser Gly Leu Asp Asp Asn Trp Asp Asp
Ala Glu Gly Tyr Tyr Ser Tyr 420 425
430 Gln Leu Gly Glu Leu Leu Asp Asp Arg Tyr Glu Ile Met Ala
Thr His 435 440 445
Gly Lys Gly Val Phe Ser Thr Val Val Arg Ala Lys Asp Thr Lys Ala 450
455 460 Glu Leu Gly Glu Pro
Glu Glu Val Ala Ile Lys Ile Ile Arg Asn Asn 465 470
475 480 Glu Thr Met His Lys Ala Gly Gln Thr Glu
Ile Gln Ile Leu Lys Lys 485 490
495 Leu Ala Gly Ser Asp Pro Glu Asn Lys Arg His Cys Val Arg Phe
Leu 500 505 510 Ser
Thr Phe Lys Tyr Arg Asn His Leu Cys Leu Val Phe Glu Ser Leu 515
520 525 His Leu Asn Leu Arg Glu
Ile Val Lys Lys Tyr Gly Arg Asn Ile Gly 530 535
540 Ile Gln Leu Ser Gly Val Arg Val Tyr Ala Thr
Gln Leu Phe Ile Ser 545 550 555
560 Leu Lys His Leu Lys Asn Cys Gly Val Leu His Cys Asp Ile Lys Pro
565 570 575 Asp Asn
Met Leu Val Asn Glu Gly Arg Asn Thr Leu Lys Leu Cys Asp 580
585 590 Phe Gly Ser Ala Met Phe Ala
Gly Thr Asn Glu Val Thr Pro Tyr Leu 595 600
605 Val Ser Arg Phe Tyr Arg Ala Pro Glu Ile Ile Leu
Gly Leu Pro Tyr 610 615 620
Asp His Pro Leu Asp Ile Trp Ser Val Gly Cys Cys Leu Tyr Glu Leu 625
630 635 640 Phe Ser Gly
Lys Ile Met Phe Pro Gly Ser Thr Asn Asn Glu Met Leu 645
650 655 Arg Leu His Met Glu Leu Lys Gly
Ala Phe Pro Lys Lys Met Leu Arg 660 665
670 Lys Gly Ala Phe Ile Asp Gln His Phe Asp Lys Asp Leu
Cys Phe Tyr 675 680 685
Ala Thr Glu Glu Asp Ser Val Thr Arg Lys Thr Thr Lys Arg Met Met 690
695 700 Val Asn Ile Lys
Pro Lys Glu Phe Gly Ser Val Ile Lys Gln Arg Tyr 705 710
715 720 Lys Asp Glu Asp Ser Lys Leu Leu Val
His Phe Arg Asp Leu Leu Asp 725 730
735 Arg Ile Phe Ile Leu Asp Pro Gln Lys Arg Ile Thr Val Ser
Gln Ala 740 745 750
Leu Ala His Pro Phe Ile Thr Gly Lys 755 760
132862DNAzea mays 13atggctgccg gcggagagct ctccccctcc cccgtgcccc
ccgcatcccc catcaagcat 60tctcgctcac ccgacgatgc ccagcccgac gcatccccga
agcgtcgcaa gcgccaccat 120caccgccgcc accaccacca ccgtcggcac cgtcacgccg
attccccggt acccgtggca 180gccgacgaag aggtagagga gggggagata ctcgaagatg
ccaccgccgc ctctgccatg 240gaggtcgatg ctgagtccgc cgccccagag gctttccttg
ctccggagca ctttggtaat 300ggtgctgata cagactcaaa cacagatgca accaagatgc
aagctcctgc tctgcctacc 360cttccatcct cagaagatgg aaggaggtca cttcatgatg
cccctgagtc tgaaagagga 420gatattcttt caagtgatgt tgaggacaac aaaggacatg
aacgaaggca aagacaaaac 480cgttctaaat ctccaaagtc tagaagggag aaggaaagga
ggcacaaaga tgaccatcac 540acttcatctt caaaagatta tcattccaga aatcactcta
gaacatctcc ttactcaagg 600catcaaagtg aagctcattc gagagatctg tacttgagat
atagagagaa aggtgattac 660actaatggtt ctcgtgcaaa tcttagggat ggttctgatc
atgagagcaa tgatcggaat 720gggaagtctg gcagacatac gactaggacc cacgggagtg
aaagagaaag gagcagcagc 780catggtattc atgatagaca tggtgacagg tacagtgaca
gacgtgccag ccaggaaagg 840catagagacg ataggatata tagagacaaa atcgattcat
tagaagctgc tcctaggcac 900agagaaagaa gcaggagtca tagtagatcg gatccaaggg
aaaatacacg tcttcgtgat 960caaagcaggg agagggaaag acggagtggt agttcaaggc
atagggatca tgacagcaag 1020agggatacaa gtaaagatcg gcatagagaa tctgacaggg
ttaacagtgc acatgaaagg 1080gatagaggga gagaggctag ggacagggaa tggcataggg
tcaagggaag tgaaactcat 1140agagctaagg aaggacggga caaagttagc gataatgata
ggcacaggga ttcaacacgc 1200tcaaaatata gcgtgtctga tggttacaaa gagaggacaa
gatctgggga gaaaggtaga 1260gatgctgacc ataaaaaccg gaagtttgaa gaaatgaagg
aaaattctct caaggaggaa 1320gatgaagagg agtaccaaga gaaaatagaa cagcagttag
caatgcagga ggaagacgac 1380cctgaaaaaa tcaaagagga agcaaggagg aggaaagaag
ctatcatggc aaagtacagg 1440cagcagcaat cgcagaaaga ggatatggaa tctaaaccaa
gtggcaatga tgaagaagta 1500agagcaatgg atggaaacga agctatacat cagaaagatg
atatcgatag cagctttatg 1560ggcaatgtcg aagctgaaaa taagcatgat tcttcagagg
tatttgatgg caagacaggc 1620tttaatgtgg gaaggtctcc tgctcacaat tatgcttcaa
ctagcacggg agcattcact 1680gatgagagga caataggtgt ttcaggtctt ggagagggtt
ctcccaagag tgagagatca 1740gcagacatgt tttgtgatga cattttcgga gaatcaccca
ccggaattag aaaattggga 1800aaggatgatg gtttgcatat tgagagaaat gctcttcatg
acaactggga tgatgcagat 1860gggtactaca cttatcggtt cggagaattg ctggatggcc
gttatgaaat catagcagca 1920catgggaagg gtgtgttctc aacagttgtg cgggcaaaag
atcttaaagc gagtaaggat 1980gatcctgaag aagttgccat caaaattatt cgcaacaatg
agacaatgta caaggctggt 2040aagcaagagg tttcaatatt ggaaaaactt gcaagtgcgg
accgtgagga caaacgccac 2100tgcgtgcggt ttatttctag tttcatgtac cggaaccatc
tttgcttagt ttttgaatct 2160ctcaatatga atcttcgtga ggttttaaag aaatttggtc
gtaatattgg gcttaaactg 2220actgcggtga gggcatattc aaagcagctt ttcatcgccc
tgaagcacct gaagaactgc 2280aaagttttgc actgtgatat aaaaccagat aatatgctgg
tgaatgaggc taagaatgtg 2340ctcaaggtat gtgattttgg caacgctatg cttgctggta
tgaatgaagt tacgccttat 2400cttgtcagcc gtttctatcg ggcacctgag atcgttcttg
ggttagccta tgatcaccct 2460ttagacatgt ggtcagttgg ttgctgtcta tatgagcttt
ataccgggaa agtcttattt 2520cctggtccat caaacaatgc catgcttcgg cttcatatgg
aattgaaggg tccattccct 2580aagaagatgc ttcgaaaggg tgcctttact atgcaacact
tcgatcaaga tctcaatttt 2640catgctaccg aggaggatcc tgtgacaaaa acggctgtga
gaaggttaat tttgaacatt 2700aaaccaaagg atgttggttc tttgtttccg aactttcctg
gcgaggatcc aaaaatgcta 2760tccagtttta aggatcttct tgataaaata tttacattag
atccagaaaa gaggataact 2820gtatcgcaag cacttagcca tccatttatc actggcaagt
ga 286214953PRTzea mays 14Met Ala Ala Gly Gly Glu
Leu Ser Pro Ser Pro Val Pro Pro Ala Ser 1 5
10 15 Pro Ile Lys His Ser Arg Ser Pro Asp Asp Ala
Gln Pro Asp Ala Ser 20 25
30 Pro Lys Arg Arg Lys Arg His His His Arg Arg His His His His
Arg 35 40 45 Arg
His Arg His Ala Asp Ser Pro Val Pro Val Ala Ala Asp Glu Glu 50
55 60 Val Glu Glu Gly Glu Ile
Leu Glu Asp Ala Thr Ala Ala Ser Ala Met 65 70
75 80 Glu Val Asp Ala Glu Ser Ala Ala Pro Glu Ala
Phe Leu Ala Pro Glu 85 90
95 His Phe Gly Asn Gly Ala Asp Thr Asp Ser Asn Thr Asp Ala Thr Lys
100 105 110 Met Gln
Ala Pro Ala Leu Pro Thr Leu Pro Ser Ser Glu Asp Gly Arg 115
120 125 Arg Ser Leu His Asp Ala Pro
Glu Ser Glu Arg Gly Asp Ile Leu Ser 130 135
140 Ser Asp Val Glu Asp Asn Lys Gly His Glu Arg Arg
Gln Arg Gln Asn 145 150 155
160 Arg Ser Lys Ser Pro Lys Ser Arg Arg Glu Lys Glu Arg Arg His Lys
165 170 175 Asp Asp His
His Thr Ser Ser Ser Lys Asp Tyr His Ser Arg Asn His 180
185 190 Ser Arg Thr Ser Pro Tyr Ser Arg
His Gln Ser Glu Ala His Ser Arg 195 200
205 Asp Leu Tyr Leu Arg Tyr Arg Glu Lys Gly Asp Tyr Thr
Asn Gly Ser 210 215 220
Arg Ala Asn Leu Arg Asp Gly Ser Asp His Glu Ser Asn Asp Arg Asn 225
230 235 240 Gly Lys Ser Gly
Arg His Thr Thr Arg Thr His Gly Ser Glu Arg Glu 245
250 255 Arg Ser Ser Ser His Gly Ile His Asp
Arg His Gly Asp Arg Tyr Ser 260 265
270 Asp Arg Arg Ala Ser Gln Glu Arg His Arg Asp Asp Arg Ile
Tyr Arg 275 280 285
Asp Lys Ile Asp Ser Leu Glu Ala Ala Pro Arg His Arg Glu Arg Ser 290
295 300 Arg Ser His Ser Arg
Ser Asp Pro Arg Glu Asn Thr Arg Leu Arg Asp 305 310
315 320 Gln Ser Arg Glu Arg Glu Arg Arg Ser Gly
Ser Ser Arg His Arg Asp 325 330
335 His Asp Ser Lys Arg Asp Thr Ser Lys Asp Arg His Arg Glu Ser
Asp 340 345 350 Arg
Val Asn Ser Ala His Glu Arg Asp Arg Gly Arg Glu Ala Arg Asp 355
360 365 Arg Glu Trp His Arg Val
Lys Gly Ser Glu Thr His Arg Ala Lys Glu 370 375
380 Gly Arg Asp Lys Val Ser Asp Asn Asp Arg His
Arg Asp Ser Thr Arg 385 390 395
400 Ser Lys Tyr Ser Val Ser Asp Gly Tyr Lys Glu Arg Thr Arg Ser Gly
405 410 415 Glu Lys
Gly Arg Asp Ala Asp His Lys Asn Arg Lys Phe Glu Glu Met 420
425 430 Lys Glu Asn Ser Leu Lys Glu
Glu Asp Glu Glu Glu Tyr Gln Glu Lys 435 440
445 Ile Glu Gln Gln Leu Ala Met Gln Glu Glu Asp Asp
Pro Glu Lys Ile 450 455 460
Lys Glu Glu Ala Arg Arg Arg Lys Glu Ala Ile Met Ala Lys Tyr Arg 465
470 475 480 Gln Gln Gln
Ser Gln Lys Glu Asp Met Glu Ser Lys Pro Ser Gly Asn 485
490 495 Asp Glu Glu Val Arg Ala Met Asp
Gly Asn Glu Ala Ile His Gln Lys 500 505
510 Asp Asp Ile Asp Ser Ser Phe Met Gly Asn Val Glu Ala
Glu Asn Lys 515 520 525
His Asp Ser Ser Glu Val Phe Asp Gly Lys Thr Gly Phe Asn Val Gly 530
535 540 Arg Ser Pro Ala
His Asn Tyr Ala Ser Thr Ser Thr Gly Ala Phe Thr 545 550
555 560 Asp Glu Arg Thr Ile Gly Val Ser Gly
Leu Gly Glu Gly Ser Pro Lys 565 570
575 Ser Glu Arg Ser Ala Asp Met Phe Cys Asp Asp Ile Phe Gly
Glu Ser 580 585 590
Pro Thr Gly Ile Arg Lys Leu Gly Lys Asp Asp Gly Leu His Ile Glu
595 600 605 Arg Asn Ala Leu
His Asp Asn Trp Asp Asp Ala Asp Gly Tyr Tyr Thr 610
615 620 Tyr Arg Phe Gly Glu Leu Leu Asp
Gly Arg Tyr Glu Ile Ile Ala Ala 625 630
635 640 His Gly Lys Gly Val Phe Ser Thr Val Val Arg Ala
Lys Asp Leu Lys 645 650
655 Ala Ser Lys Asp Asp Pro Glu Glu Val Ala Ile Lys Ile Ile Arg Asn
660 665 670 Asn Glu Thr
Met Tyr Lys Ala Gly Lys Gln Glu Val Ser Ile Leu Glu 675
680 685 Lys Leu Ala Ser Ala Asp Arg Glu
Asp Lys Arg His Cys Val Arg Phe 690 695
700 Ile Ser Ser Phe Met Tyr Arg Asn His Leu Cys Leu Val
Phe Glu Ser 705 710 715
720 Leu Asn Met Asn Leu Arg Glu Val Leu Lys Lys Phe Gly Arg Asn Ile
725 730 735 Gly Leu Lys Leu
Thr Ala Val Arg Ala Tyr Ser Lys Gln Leu Phe Ile 740
745 750 Ala Leu Lys His Leu Lys Asn Cys Lys
Val Leu His Cys Asp Ile Lys 755 760
765 Pro Asp Asn Met Leu Val Asn Glu Ala Lys Asn Val Leu Lys
Val Cys 770 775 780
Asp Phe Gly Asn Ala Met Leu Ala Gly Met Asn Glu Val Thr Pro Tyr 785
790 795 800 Leu Val Ser Arg Phe
Tyr Arg Ala Pro Glu Ile Val Leu Gly Leu Ala 805
810 815 Tyr Asp His Pro Leu Asp Met Trp Ser Val
Gly Cys Cys Leu Tyr Glu 820 825
830 Leu Tyr Thr Gly Lys Val Leu Phe Pro Gly Pro Ser Asn Asn Ala
Met 835 840 845 Leu
Arg Leu His Met Glu Leu Lys Gly Pro Phe Pro Lys Lys Met Leu 850
855 860 Arg Lys Gly Ala Phe Thr
Met Gln His Phe Asp Gln Asp Leu Asn Phe 865 870
875 880 His Ala Thr Glu Glu Asp Pro Val Thr Lys Thr
Ala Val Arg Arg Leu 885 890
895 Ile Leu Asn Ile Lys Pro Lys Asp Val Gly Ser Leu Phe Pro Asn Phe
900 905 910 Pro Gly
Glu Asp Pro Lys Met Leu Ser Ser Phe Lys Asp Leu Leu Asp 915
920 925 Lys Ile Phe Thr Leu Asp Pro
Glu Lys Arg Ile Thr Val Ser Gln Ala 930 935
940 Leu Ser His Pro Phe Ile Thr Gly Lys 945
950 152784DNAglycine max 15atggccaccg atgcccgtga
ctcgcgtcgc aagcatcacc gatcttcctc tccagaggac 60gtggacagat cctcgaagcg
tcacaagcac cgccataaca gccatcgcca ccgccacggg 120agcaaaaagc gcgacgaaga
ggttgaattc gatgatcgaa caattgctgc ggttccttct 180ccaacttcgc acagatatct
ccatgacgac gatgtggagg agggagagat ccttgaagat 240gaggctctcg acggtgaggt
tggaaaaaag gagacggaat ctgatgttga acccggtgaa 300atcaaggtga caggagatcg
agatgttcga tctgataatc aaaattcgga acccctcact 360aaaatttcag aaactagaaa
tgaagacatt agggatgata aatttattag tcctgcaatt 420gatgcacaag atgatgtctc
tcctaatcgt tcaagttctg agactcgaga tggaaagcat 480gctcaagctc gtacagatgg
tgtgggcaat ggttatttgg atcctaaatc ttccaaaggg 540gataagtggc agaatgggga
acttggacat tttaaagggg aaaagtatag aatgtcagga 600agttctcctt ctcatggtag
atatagaagt cgatcaagat caattggtca tactagagac 660aggtctcgct ctcgtagtat
catagacgaa tatcctcatt ctaagagaag gcgctttgac 720tatgaccatg atgaggaaag
agtgagggca cgtggaaggg agcatgaggc cagggataga 780gatgtggata gagacttgca
cggagaaaag aagcaagagg aaactagcag gggtaaagag 840attgagagcc gtgataggta
tagaagggat atagaaaaag ataggagcag ggagagggag 900gaggataggg acaggagaca
agaaaaggaa agagatagga gctgggatac agtgatggag 960agggatagga gaagggaaaa
ggaaagagat agaagtaggg acagaataag aggtggcaag 1020agagataaag acccagagaa
tgaaagggat gataagcatc gggcaagaga taatattaag 1080aagagggaaa gacatgatga
taaatatagg cacaaagata gagacactgc taatgataga 1140tatagaaaac attcaaggca
tgaagaaaat gaatatcgtt gggaaagaaa aagaaattct 1200gataatcctg taaaggttta
tagctcaatg ggaagtactg cagaagtggg tgaaagcaaa 1260ctaacaagca gtgaggttga
accagatgac ttagaggagg atacattaca attacctgag 1320caagaagagg aagatctcaa
caggatcaaa gaagagagta gaagaagaag ggaagcaata 1380atggagaaat acaagaagca
gcatcagcaa gtagaagaag cggttggaaa tgaaggaaac 1440ggtattattt tccccatttc
attaacgatc tgtaattatt cttacaagaa ggcagccatt 1500cctaatgaca tctctgaagc
tcgtgatggt aaaaatgatg atgctgatta tttggagcca 1560tcatttgctg ttgggaaatc
tcctgaaaat gtgaatgttg cttctaagaa gatgtctcct 1620gctggaggtc tgggagaggg
tactccaaag agtgaaaggt cagaggacaa gttttgtgat 1680gatatatttg gtgagacgcc
aacgggagtt cggaaatcag gaaaaggaga tggtttactg 1740attgagaggg ctggcctaca
tgacaattgg gacgatgcag agggttatta tagctatcgt 1800attggtgaaa tacttgatgg
ccgatatgaa gtcactgctg cacatgggag gggtgtcttt 1860tcaacagttg ttcgcggaaa
gaatctaaag atgggaaatg gtgagccaga agaagtagcc 1920ataaaaatta ttcgtagtaa
tgacaccatg tacaaggctg gtatggatga attggtcata 1980ttgaagaaat tagtaggtgc
agatccagat gataagcgtc attgtgttcg tttcctttca 2040agttttagat acaggaatca
tctttgttta gtttttgaat ctctaaatat gaatctgcga 2100gaggttttaa agaagtttgg
tcgcaatatt ggccttaggc taacagctgt gagagcatac 2160gcaaaacagc tttttattgc
tctgaagcat ctccggaact gtggtgttct tcattgtgat 2220ataaagccag ataatatgtt
ggtaaatgag tctaaaaatg ttttgaagct ttgtgacttt 2280ggcaatgcca tgtttgctgg
taaaaatgaa gttacaccat atcttgtgag tcgtttttat 2340cgtgccccgg aaataatact
tggcttgcca tatgatcatc cattggatat ttggtctgta 2400ggttgttgtt tgtatgagtt
gtatataggg aaggttcttt tcccaggtct tacaaacaat 2460gacatgctac ggcttcacat
ggaactgaag ggtccttttc caaagaagat gctgcgtaag 2520ggagcattta ctgaacagca
ttttgatcag gatctgaatt ttcttgctac tgaggaggat 2580cctgtaacaa aaaagaccat
aaagcggctg atactcaaca ttaagccaaa agatattggg 2640acactcatta ctggctctcc
tggggaggat ccaaaaatgt tagccaactt taaggatctt 2700ctggaaaaag tttttgtctt
ggatccagac aagaggctga cagtgtcaca agctctgaac 2760cacccattca tcactggcaa
gtga 278416927PRTglycine max
16Met Ala Thr Asp Ala Arg Asp Ser Arg Arg Lys His His Arg Ser Ser 1
5 10 15 Ser Pro Glu Asp
Val Asp Arg Ser Ser Lys Arg His Lys His Arg His 20
25 30 Asn Ser His Arg His Arg His Gly Ser
Lys Lys Arg Asp Glu Glu Val 35 40
45 Glu Phe Asp Asp Arg Thr Ile Ala Ala Val Pro Ser Pro Thr
Ser His 50 55 60
Arg Tyr Leu His Asp Asp Asp Val Glu Glu Gly Glu Ile Leu Glu Asp 65
70 75 80 Glu Ala Leu Asp Gly
Glu Val Gly Lys Lys Glu Thr Glu Ser Asp Val 85
90 95 Glu Pro Gly Glu Ile Lys Val Thr Gly Asp
Arg Asp Val Arg Ser Asp 100 105
110 Asn Gln Asn Ser Glu Pro Leu Thr Lys Ile Ser Glu Thr Arg Asn
Glu 115 120 125 Asp
Ile Arg Asp Asp Lys Phe Ile Ser Pro Ala Ile Asp Ala Gln Asp 130
135 140 Asp Val Ser Pro Asn Arg
Ser Ser Ser Glu Thr Arg Asp Gly Lys His 145 150
155 160 Ala Gln Ala Arg Thr Asp Gly Val Gly Asn Gly
Tyr Leu Asp Pro Lys 165 170
175 Ser Ser Lys Gly Asp Lys Trp Gln Asn Gly Glu Leu Gly His Phe Lys
180 185 190 Gly Glu
Lys Tyr Arg Met Ser Gly Ser Ser Pro Ser His Gly Arg Tyr 195
200 205 Arg Ser Arg Ser Arg Ser Ile
Gly His Thr Arg Asp Arg Ser Arg Ser 210 215
220 Arg Ser Ile Ile Asp Glu Tyr Pro His Ser Lys Arg
Arg Arg Phe Asp 225 230 235
240 Tyr Asp His Asp Glu Glu Arg Val Arg Ala Arg Gly Arg Glu His Glu
245 250 255 Ala Arg Asp
Arg Asp Val Asp Arg Asp Leu His Gly Glu Lys Lys Gln 260
265 270 Glu Glu Thr Ser Arg Gly Lys Glu
Ile Glu Ser Arg Asp Arg Tyr Arg 275 280
285 Arg Asp Ile Glu Lys Asp Arg Ser Arg Glu Arg Glu Glu
Asp Arg Asp 290 295 300
Arg Arg Gln Glu Lys Glu Arg Asp Arg Ser Trp Asp Thr Val Met Glu 305
310 315 320 Arg Asp Arg Arg
Arg Glu Lys Glu Arg Asp Arg Ser Arg Asp Arg Ile 325
330 335 Arg Gly Gly Lys Arg Asp Lys Asp Pro
Glu Asn Glu Arg Asp Asp Lys 340 345
350 His Arg Ala Arg Asp Asn Ile Lys Lys Arg Glu Arg His Asp
Asp Lys 355 360 365
Tyr Arg His Lys Asp Arg Asp Thr Ala Asn Asp Arg Tyr Arg Lys His 370
375 380 Ser Arg His Glu Glu
Asn Glu Tyr Arg Trp Glu Arg Lys Arg Asn Ser 385 390
395 400 Asp Asn Pro Val Lys Val Tyr Ser Ser Met
Gly Ser Thr Ala Glu Val 405 410
415 Gly Glu Ser Lys Leu Thr Ser Ser Glu Val Glu Pro Asp Asp Leu
Glu 420 425 430 Glu
Asp Thr Leu Gln Leu Pro Glu Gln Glu Glu Glu Asp Leu Asn Arg 435
440 445 Ile Lys Glu Glu Ser Arg
Arg Arg Arg Glu Ala Ile Met Glu Lys Tyr 450 455
460 Lys Lys Gln His Gln Gln Val Glu Glu Ala Val
Gly Asn Glu Gly Asn 465 470 475
480 Gly Ile Ile Phe Pro Ile Ser Leu Thr Ile Cys Asn Tyr Ser Tyr Lys
485 490 495 Lys Ala
Ala Ile Pro Asn Asp Ile Ser Glu Ala Arg Asp Gly Lys Asn 500
505 510 Asp Asp Ala Asp Tyr Leu Glu
Pro Ser Phe Ala Val Gly Lys Ser Pro 515 520
525 Glu Asn Val Asn Val Ala Ser Lys Lys Met Ser Pro
Ala Gly Gly Leu 530 535 540
Gly Glu Gly Thr Pro Lys Ser Glu Arg Ser Glu Asp Lys Phe Cys Asp 545
550 555 560 Asp Ile Phe
Gly Glu Thr Pro Thr Gly Val Arg Lys Ser Gly Lys Gly 565
570 575 Asp Gly Leu Leu Ile Glu Arg Ala
Gly Leu His Asp Asn Trp Asp Asp 580 585
590 Ala Glu Gly Tyr Tyr Ser Tyr Arg Ile Gly Glu Ile Leu
Asp Gly Arg 595 600 605
Tyr Glu Val Thr Ala Ala His Gly Arg Gly Val Phe Ser Thr Val Val 610
615 620 Arg Gly Lys Asn
Leu Lys Met Gly Asn Gly Glu Pro Glu Glu Val Ala 625 630
635 640 Ile Lys Ile Ile Arg Ser Asn Asp Thr
Met Tyr Lys Ala Gly Met Asp 645 650
655 Glu Leu Val Ile Leu Lys Lys Leu Val Gly Ala Asp Pro Asp
Asp Lys 660 665 670
Arg His Cys Val Arg Phe Leu Ser Ser Phe Arg Tyr Arg Asn His Leu
675 680 685 Cys Leu Val Phe
Glu Ser Leu Asn Met Asn Leu Arg Glu Val Leu Lys 690
695 700 Lys Phe Gly Arg Asn Ile Gly Leu
Arg Leu Thr Ala Val Arg Ala Tyr 705 710
715 720 Ala Lys Gln Leu Phe Ile Ala Leu Lys His Leu Arg
Asn Cys Gly Val 725 730
735 Leu His Cys Asp Ile Lys Pro Asp Asn Met Leu Val Asn Glu Ser Lys
740 745 750 Asn Val Leu
Lys Leu Cys Asp Phe Gly Asn Ala Met Phe Ala Gly Lys 755
760 765 Asn Glu Val Thr Pro Tyr Leu Val
Ser Arg Phe Tyr Arg Ala Pro Glu 770 775
780 Ile Ile Leu Gly Leu Pro Tyr Asp His Pro Leu Asp Ile
Trp Ser Val 785 790 795
800 Gly Cys Cys Leu Tyr Glu Leu Tyr Ile Gly Lys Val Leu Phe Pro Gly
805 810 815 Leu Thr Asn Asn
Asp Met Leu Arg Leu His Met Glu Leu Lys Gly Pro 820
825 830 Phe Pro Lys Lys Met Leu Arg Lys Gly
Ala Phe Thr Glu Gln His Phe 835 840
845 Asp Gln Asp Leu Asn Phe Leu Ala Thr Glu Glu Asp Pro Val
Thr Lys 850 855 860
Lys Thr Ile Lys Arg Leu Ile Leu Asn Ile Lys Pro Lys Asp Ile Gly 865
870 875 880 Thr Leu Ile Thr Gly
Ser Pro Gly Glu Asp Pro Lys Met Leu Ala Asn 885
890 895 Phe Lys Asp Leu Leu Glu Lys Val Phe Val
Leu Asp Pro Asp Lys Arg 900 905
910 Leu Thr Val Ser Gln Ala Leu Asn His Pro Phe Ile Thr Gly Lys
915 920 925
172597DNAoryza sativa 17atggctgagg aaccctcccc ctccccctcc tcctcctgcg
ccaagcacca ccgctctccc 60gaccccgccg accccgccgc ctctccatga actcagatgc
tgatgcaacc gtgctgcatg 120cttctcgctt gcctactcat tcatcctcaa gggacgaaac
caagtcaaat cacaccgccc 180atgagcctga gagtggaggc gatgcagatg ataccaaagg
ggatagacaa agtcaaaggg 240tgccaaaatc accactattg acaagggaga aagaaaggaa
gcacaaagat gagcaccgca 300aatcgtatcc taaagattca cattccaaag agcagtctag
aagatcccct tcaaggcacc 360atagcagtca agatcatgcc aggcatcact caaggtctag
agatactggt gctgaagcta 420atggttcgcg ggcaagtaca agggaagatt ctgaccgtga
cagcaacggc agaaatagta 480agcatggtag gcatgcaacc aggagtcgag ataatgagac
agaaaggagc agcagctatg 540ctgttcgtga tgaggcgtat gatgagcggg aaagatataa
gcatgaaaga aggcatagaa 600gcaacccagt tgatagagac aaagtggatt tgcatgaact
aactcacagg gatagagaaa 660ggagcagcag tcgcagtaga tctgatcgta gggagagtgc
acatcacatt cgtgatgaaa 720gcagggagag tgaaaggcgg agtagtagtt caaggcataa
agataatgag agaagggata 780gaagtaagga tcgctataaa gaatctgaca aggttgacag
tggacatgaa agggacaaaa 840caagagatga tagagacagg ggacgacata aggatttgga
aagtagaaag cggagaaatg 900gagaagcaaa ggacagggat gacaggcaca aggattctac
acgctcaaaa tacagtactt 960ctgatagtca taaacaccgc tcaagatcca gggagagagg
tagagatgct gaacgtagag 1020gccagagatc tgaagagctg aaggagaata ctttcaggga
ggaggatgaa gaggagtacc 1080aagagaaaat tgaacagcag ttagcaatgc aggaagaaga
ggaccctgaa aaaattaagg 1140aggaagcaag gaggaggaaa gaagctatta tggcaaaata
caggcagcaa caattgcaga 1200agcagcagct ggaatcttta cctagaagta atgatgaaga
agaagtggaa atgaacagag 1260gtgataatgc agatctgaaa ggtgataacg atagcagatt
tgtggctagc gaggaagctg 1320aaaataagca tgattcttca gatgcaattg ttggtgaaac
agacttcact gtgggaaagt 1380ctcctgctca caatgatggt gcaggaactt tgggtaatca
gagaacaact ggtgtttcag 1440gtcttggaga gggcactcca aagagtgaga gatcggcgga
tatgttttgt gatgacattt 1500ttggagaatc acctgctggc atccggaaat tgggcaagga
tgatggtttg cgcatcgaga 1560aaaatgctct tcatgacaac tgggatgatg ccgaggggta
ctatacttac cgttttgggg 1620aattgctgga tgggcgctat gagattacag cagcacatgg
aaagggagtg ttttcaacag 1680ttgtccgagc aaaagatctt aaagctggga aggatgatcc
cgaagaggtt gctattaaga 1740ttattcgcaa caatgagaca atgtacaagg ctggtaagca
agaggtttca atattagaaa 1800aactggcaag tgcggatcgt gaagacaggc gccactgcgt
gcggtttatt tctagtttca 1860tgtataggaa ccatctttgc ttagtttttg aatctctaaa
tatgaatctt cgtgaggtac 1920taaagaaatt tggtcgcaat atcggactta aactaactgc
tgtgagggca tattcaaagc 1980agcttttcat cgccctgaag catctgaaaa actgcaaagt
gctgcactgt gatataaagc 2040cagataatat gctggtgaat gaggctaaga atgtgctgaa
gctctgtgat tttggcaatg 2100caatgcttgc tggaatgaac gaggttacac cttatcttgt
gagccgtttc tatcgtgcac 2160ctgagataat tcttgggtta ccctacgacc acccattaga
catgtggtca gttggctgct 2220gtctatatga actttacacc ggaaaagtcc tatttccagg
tccatcaaat aatgacatgc 2280ttcggcttca tatggaactg aagggcccct tccccaagaa
aatgcttcga aagggtgcct 2340ttacgatgca acattttgac caagatctca actttcatgc
cactgaggag gatcctgtga 2400ctaaaaaggc tgtgacaagg atgattttga acattaagcc
aaaggatatt ggttccttga 2460tttcaaactt ccctggcgag gatccaaaaa tgctatccaa
ctttaaagat cttcttgaaa 2520aaatatttgt cttagatcca gaaaagagga taaccatatc
acaagcactt agccatccat 2580ttatcactgg caagtga
259718836PRToryza sativa 18Met Asn Ser Asp Ala Asp
Ala Thr Val Leu His Ala Ser Arg Leu Pro 1 5
10 15 Thr His Ser Ser Ser Arg Asp Glu Thr Lys Ser
Asn His Thr Ala His 20 25
30 Glu Pro Glu Ser Gly Gly Asp Ala Asp Asp Thr Lys Gly Asp Arg
Gln 35 40 45 Ser
Gln Arg Val Pro Lys Ser Pro Leu Leu Thr Arg Glu Lys Glu Arg 50
55 60 Lys His Lys Asp Glu His
Arg Lys Ser Tyr Pro Lys Asp Ser His Ser 65 70
75 80 Lys Glu Gln Ser Arg Arg Ser Pro Ser Arg His
His Ser Ser Gln Asp 85 90
95 His Ala Arg His His Ser Arg Ser Arg Asp Thr Gly Ala Glu Ala Asn
100 105 110 Gly Ser
Arg Ala Ser Thr Arg Glu Asp Ser Asp Arg Asp Ser Asn Gly 115
120 125 Arg Asn Ser Lys His Gly Arg
His Ala Thr Arg Ser Arg Asp Asn Glu 130 135
140 Thr Glu Arg Ser Ser Ser Tyr Ala Val Arg Asp Glu
Ala Tyr Asp Glu 145 150 155
160 Arg Glu Arg Tyr Lys His Glu Arg Arg His Arg Ser Asn Pro Val Asp
165 170 175 Arg Asp Lys
Val Asp Leu His Glu Leu Thr His Arg Asp Arg Glu Arg 180
185 190 Ser Ser Ser Arg Ser Arg Ser Asp
Arg Arg Glu Ser Ala His His Ile 195 200
205 Arg Asp Glu Ser Arg Glu Ser Glu Arg Arg Ser Ser Ser
Ser Arg His 210 215 220
Lys Asp Asn Glu Arg Arg Asp Arg Ser Lys Asp Arg Tyr Lys Glu Ser 225
230 235 240 Asp Lys Val Asp
Ser Gly His Glu Arg Asp Lys Thr Arg Asp Asp Arg 245
250 255 Asp Arg Gly Arg His Lys Asp Leu Glu
Ser Arg Lys Arg Arg Asn Gly 260 265
270 Glu Ala Lys Asp Arg Asp Asp Arg His Lys Asp Ser Thr Arg
Ser Lys 275 280 285
Tyr Ser Thr Ser Asp Ser His Lys His Arg Ser Arg Ser Arg Glu Arg 290
295 300 Gly Arg Asp Ala Glu
Arg Arg Gly Gln Arg Ser Glu Glu Leu Lys Glu 305 310
315 320 Asn Thr Phe Arg Glu Glu Asp Glu Glu Glu
Tyr Gln Glu Lys Ile Glu 325 330
335 Gln Gln Leu Ala Met Gln Glu Glu Glu Asp Pro Glu Lys Ile Lys
Glu 340 345 350 Glu
Ala Arg Arg Arg Lys Glu Ala Ile Met Ala Lys Tyr Arg Gln Gln 355
360 365 Gln Leu Gln Lys Gln Gln
Leu Glu Ser Leu Pro Arg Ser Asn Asp Glu 370 375
380 Glu Glu Val Glu Met Asn Arg Gly Asp Asn Ala
Asp Leu Lys Gly Asp 385 390 395
400 Asn Asp Ser Arg Phe Val Ala Ser Glu Glu Ala Glu Asn Lys His Asp
405 410 415 Ser Ser
Asp Ala Ile Val Gly Glu Thr Asp Phe Thr Val Gly Lys Ser 420
425 430 Pro Ala His Asn Asp Gly Ala
Gly Thr Leu Gly Asn Gln Arg Thr Thr 435 440
445 Gly Val Ser Gly Leu Gly Glu Gly Thr Pro Lys Ser
Glu Arg Ser Ala 450 455 460
Asp Met Phe Cys Asp Asp Ile Phe Gly Glu Ser Pro Ala Gly Ile Arg 465
470 475 480 Lys Leu Gly
Lys Asp Asp Gly Leu Arg Ile Glu Lys Asn Ala Leu His 485
490 495 Asp Asn Trp Asp Asp Ala Glu Gly
Tyr Tyr Thr Tyr Arg Phe Gly Glu 500 505
510 Leu Leu Asp Gly Arg Tyr Glu Ile Thr Ala Ala His Gly
Lys Gly Val 515 520 525
Phe Ser Thr Val Val Arg Ala Lys Asp Leu Lys Ala Gly Lys Asp Asp 530
535 540 Pro Glu Glu Val
Ala Ile Lys Ile Ile Arg Asn Asn Glu Thr Met Tyr 545 550
555 560 Lys Ala Gly Lys Gln Glu Val Ser Ile
Leu Glu Lys Leu Ala Ser Ala 565 570
575 Asp Arg Glu Asp Arg Arg His Cys Val Arg Phe Ile Ser Ser
Phe Met 580 585 590
Tyr Arg Asn His Leu Cys Leu Val Phe Glu Ser Leu Asn Met Asn Leu
595 600 605 Arg Glu Val Leu
Lys Lys Phe Gly Arg Asn Ile Gly Leu Lys Leu Thr 610
615 620 Ala Val Arg Ala Tyr Ser Lys Gln
Leu Phe Ile Ala Leu Lys His Leu 625 630
635 640 Lys Asn Cys Lys Val Leu His Cys Asp Ile Lys Pro
Asp Asn Met Leu 645 650
655 Val Asn Glu Ala Lys Asn Val Leu Lys Leu Cys Asp Phe Gly Asn Ala
660 665 670 Met Leu Ala
Gly Met Asn Glu Val Thr Pro Tyr Leu Val Ser Arg Phe 675
680 685 Tyr Arg Ala Pro Glu Ile Ile Leu
Gly Leu Pro Tyr Asp His Pro Leu 690 695
700 Asp Met Trp Ser Val Gly Cys Cys Leu Tyr Glu Leu Tyr
Thr Gly Lys 705 710 715
720 Val Leu Phe Pro Gly Pro Ser Asn Asn Asp Met Leu Arg Leu His Met
725 730 735 Glu Leu Lys Gly
Pro Phe Pro Lys Lys Met Leu Arg Lys Gly Ala Phe 740
745 750 Thr Met Gln His Phe Asp Gln Asp Leu
Asn Phe His Ala Thr Glu Glu 755 760
765 Asp Pro Val Thr Lys Lys Ala Val Thr Arg Met Ile Leu Asn
Ile Lys 770 775 780
Pro Lys Asp Ile Gly Ser Leu Ile Ser Asn Phe Pro Gly Glu Asp Pro 785
790 795 800 Lys Met Leu Ser Asn
Phe Lys Asp Leu Leu Glu Lys Ile Phe Val Leu 805
810 815 Asp Pro Glu Lys Arg Ile Thr Ile Ser Gln
Ala Leu Ser His Pro Phe 820 825
830 Ile Thr Gly Lys 835 191134DNAgossypium hirsutum
19ggcacgaggc cggttataag gagtggtctt catgacaatt gggatgacgc tgaaggatat
60tatagctatc gatttggtga aatacttgat ggccgatatg aagtaactgc tgctcatgga
120aaaggagttt tttcaacggt tgtacgtgcg aaggatctta aggctggtgc tactgggggg
180gaagaagtag ctataaagat cattcgtaat aatgaaacga tgcacaaggc tggtcagctg
240gaggttcaaa tattgaaaaa attagcaggt gcagatccag atgataagcg acattgtgtt
300cgttttttgt caagtttcaa gtacaggaat catctttgtt tagtttttga gtctcttcat
360atgaatctgc gtgaagttct caagaagttt ggtcgcaata ttggtcttaa actaactgct
420gtcagggctt atgctaagca actttttatt gcgcttaagc atctaaaaaa ctgtggtgtt
480cttcattgtg atataaagcc tgataacatg ctggtaaatg aggcaaaaaa tgtgctgaag
540ctttgtgatt ttggtaatgc catgtttgct ggtaaaaatg aaattacacc ataccttgtt
600agccgctttt atcgtgcacc agaaattatt cttggtttgc cttacgatca tccaatggat
660atctggtctg ttggttgctg tttgtatgag ctatatactg gaaaagttct tttccctggt
720ccaacaaaca atgacatgct acgtcttcat atggaactca aaggtccttt tccaaagaag
780atgttgcgta agggagcatt tacagaacaa cactttgatc aagatctgaa ttttcatgct
840acagaagagg atcctgttac taaaaagagt ataaagagga tgattcttaa cataaagcca
900aaagatatca gttcaattat tgttggctct ccaggtgagg atccaaagat ggtagccaac
960ttcaaagacc ttctagaaaa aatttttgtg cttgatccag agaagagaat gacagttact
1020caggcattgg ctcatccatt catcacgggc aagtggaaac atgttgctga ttttatgtct
1080ccagaaatgt gttgcgctag atatttgtac attatgaccc taactcatat ttag
113420377PRTgossupium hirsutum 20Gly Thr Arg Pro Val Ile Arg Ser Gly Leu
His Asp Asn Trp Asp Asp 1 5 10
15 Ala Glu Gly Tyr Tyr Ser Tyr Arg Phe Gly Glu Ile Leu Asp Gly
Arg 20 25 30 Tyr
Glu Val Thr Ala Ala His Gly Lys Gly Val Phe Ser Thr Val Val 35
40 45 Arg Ala Lys Asp Leu Lys
Ala Gly Ala Thr Gly Gly Glu Glu Val Ala 50 55
60 Ile Lys Ile Ile Arg Asn Asn Glu Thr Met His
Lys Ala Gly Gln Leu 65 70 75
80 Glu Val Gln Ile Leu Lys Lys Leu Ala Gly Ala Asp Pro Asp Asp Lys
85 90 95 Arg His
Cys Val Arg Phe Leu Ser Ser Phe Lys Tyr Arg Asn His Leu 100
105 110 Cys Leu Val Phe Glu Ser Leu
His Met Asn Leu Arg Glu Val Leu Lys 115 120
125 Lys Phe Gly Arg Asn Ile Gly Leu Lys Leu Thr Ala
Val Arg Ala Tyr 130 135 140
Ala Lys Gln Leu Phe Ile Ala Leu Lys His Leu Lys Asn Cys Gly Val 145
150 155 160 Leu His Cys
Asp Ile Lys Pro Asp Asn Met Leu Val Asn Glu Ala Lys 165
170 175 Asn Val Leu Lys Leu Cys Asp Phe
Gly Asn Ala Met Phe Ala Gly Lys 180 185
190 Asn Glu Ile Thr Pro Tyr Leu Val Ser Arg Phe Tyr Arg
Ala Pro Glu 195 200 205
Ile Ile Leu Gly Leu Pro Tyr Asp His Pro Met Asp Ile Trp Ser Val 210
215 220 Gly Cys Cys Leu
Tyr Glu Leu Tyr Thr Gly Lys Val Leu Phe Pro Gly 225 230
235 240 Pro Thr Asn Asn Asp Met Leu Arg Leu
His Met Glu Leu Lys Gly Pro 245 250
255 Phe Pro Lys Lys Met Leu Arg Lys Gly Ala Phe Thr Glu Gln
His Phe 260 265 270
Asp Gln Asp Leu Asn Phe His Ala Thr Glu Glu Asp Pro Val Thr Lys
275 280 285 Lys Ser Ile Lys
Arg Met Ile Leu Asn Ile Lys Pro Lys Asp Ile Ser 290
295 300 Ser Ile Ile Val Gly Ser Pro Gly
Glu Asp Pro Lys Met Val Ala Asn 305 310
315 320 Phe Lys Asp Leu Leu Glu Lys Ile Phe Val Leu Asp
Pro Glu Lys Arg 325 330
335 Met Thr Val Thr Gln Ala Leu Ala His Pro Phe Ile Thr Gly Lys Trp
340 345 350 Lys His Val
Ala Asp Phe Met Ser Pro Glu Met Cys Cys Ala Arg Tyr 355
360 365 Leu Tyr Ile Met Thr Leu Thr His
Ile 370 375 211413DNAArabidopsis thaliana
21atgggttatc tctcttgcaa cggcgaatcc gccgtcgcaa tctgcgatac ttataactgg
60aatcctcgtc gacgatctaa agtaccggag aaacgtcatc ctcctaagct tcgggttttc
120aactacgacg aactcgccgt cgctactaac ggcttctccg ccaataactt cctcggaaaa
180ggaagtcacg gcagagttta caaagcagtt ctcgacgacg gaaagcttct cgccgccgtc
240aagagaacaa caatcaccac taccgttggt aataacaata acaacgtgag tcaggtagac
300aatgagatcg agattctttc acgggttcgt caccgttgga tggtcaactt aatcggttac
360tgcgttgacc accggaggaa aacaaagctg ttagtcgtcg agtacatgcc taacggtacg
420cttcacgatc agttacattc tcgtagttcg ttagattcac ggttaagtag ttggaatcgg
480agaattaaac acgcgcttca gatcgcgatt gctgtccacg ctcttcacac cgcagagact
540caagtgatcc accgtgacat taaatcgtgt aacgttttaa tagacggtga cggtaacgct
600aggttagctg atttcggatt agcattgatc ggaaacgttg acgatgagcg tttgaaatat
660actccgccgg cgggtacgtt gggatattta gatccgtcgt acttagcacc ggcggacttg
720acggctaaga gcgatgtttt cagctttggg atattgttgt tggagattat tagtggtaga
780gaagccattg atttgaatta tagtccgtcg tgtatcgttg attgggcggt gccgcttatc
840aaacgcggcg attacgacgc gatttgtgat ttgaagatta agaaccgtcc ttattacgcc
900gtgattcgga agttggccgt tatggcggct aggtgtgtta gatcgacggc gaagaaacgt
960ccagatatgt tagaggttgt tgagtgtttg aaaacggtga ggcagttatc tccggcgtgg
1020aataaactgc ggcggaggag tgaagagaga tcggaaaatg ttttggcggt tgaggaagag
1080aaggaagaga ttcatgtgag gattgtgaga ggaggaagca ggaagaatcg gaaggtatcg
1140aacgtgacga cgagtgtgga tgatgtttac gagagattag ttccggagga aacgctgccg
1200tttcgtcgtc ggaattttgt gctgagatcg agatcagtag gagcgaaagt tggaccggat
1260ccatacgacg ggtttggtga tgagacggtg gttacaatga gattacttat tgagaaagaa
1320agaccggtga cgacggcagc gatgaggctg agtaagtcga ggtcggtggg gattgtacgt
1380agtcataaaa cggcgtcgcg gaagagatac tga
141322470PRTArabidopsis thaliana 22Met Gly Tyr Leu Ser Cys Asn Gly Glu
Ser Ala Val Ala Ile Cys Asp 1 5 10
15 Thr Tyr Asn Trp Asn Pro Arg Arg Arg Ser Lys Val Pro Glu
Lys Arg 20 25 30
His Pro Pro Lys Leu Arg Val Phe Asn Tyr Asp Glu Leu Ala Val Ala
35 40 45 Thr Asn Gly Phe
Ser Ala Asn Asn Phe Leu Gly Lys Gly Ser His Gly 50
55 60 Arg Val Tyr Lys Ala Val Leu Asp
Asp Gly Lys Leu Leu Ala Ala Val 65 70
75 80 Lys Arg Thr Thr Ile Thr Thr Thr Val Gly Asn Asn
Asn Asn Asn Val 85 90
95 Ser Gln Val Asp Asn Glu Ile Glu Ile Leu Ser Arg Val Arg His Arg
100 105 110 Trp Met Val
Asn Leu Ile Gly Tyr Cys Val Asp His Arg Arg Lys Thr 115
120 125 Lys Leu Leu Val Val Glu Tyr Met
Pro Asn Gly Thr Leu His Asp Gln 130 135
140 Leu His Ser Arg Ser Ser Leu Asp Ser Arg Leu Ser Ser
Trp Asn Arg 145 150 155
160 Arg Ile Lys His Ala Leu Gln Ile Ala Ile Ala Val His Ala Leu His
165 170 175 Thr Ala Glu Thr
Gln Val Ile His Arg Asp Ile Lys Ser Cys Asn Val 180
185 190 Leu Ile Asp Gly Asp Gly Asn Ala Arg
Leu Ala Asp Phe Gly Leu Ala 195 200
205 Leu Ile Gly Asn Val Asp Asp Glu Arg Leu Lys Tyr Thr Pro
Pro Ala 210 215 220
Gly Thr Leu Gly Tyr Leu Asp Pro Ser Tyr Leu Ala Pro Ala Asp Leu 225
230 235 240 Thr Ala Lys Ser Asp
Val Phe Ser Phe Gly Ile Leu Leu Leu Glu Ile 245
250 255 Ile Ser Gly Arg Glu Ala Ile Asp Leu Asn
Tyr Ser Pro Ser Cys Ile 260 265
270 Val Asp Trp Ala Val Pro Leu Ile Lys Arg Gly Asp Tyr Asp Ala
Ile 275 280 285 Cys
Asp Leu Lys Ile Lys Asn Arg Pro Tyr Tyr Ala Val Ile Arg Lys 290
295 300 Leu Ala Val Met Ala Ala
Arg Cys Val Arg Ser Thr Ala Lys Lys Arg 305 310
315 320 Pro Asp Met Leu Glu Val Val Glu Cys Leu Lys
Thr Val Arg Gln Leu 325 330
335 Ser Pro Ala Trp Asn Lys Leu Arg Arg Arg Ser Glu Glu Arg Ser Glu
340 345 350 Asn Val
Leu Ala Val Glu Glu Glu Lys Glu Glu Ile His Val Arg Ile 355
360 365 Val Arg Gly Gly Ser Arg Lys
Asn Arg Lys Val Ser Asn Val Thr Thr 370 375
380 Ser Val Asp Asp Val Tyr Glu Arg Leu Val Pro Glu
Glu Thr Leu Pro 385 390 395
400 Phe Arg Arg Arg Asn Phe Val Leu Arg Ser Arg Ser Val Gly Ala Lys
405 410 415 Val Gly Pro
Asp Pro Tyr Asp Gly Phe Gly Asp Glu Thr Val Val Thr 420
425 430 Met Arg Leu Leu Ile Glu Lys Glu
Arg Pro Val Thr Thr Ala Ala Met 435 440
445 Arg Leu Ser Lys Ser Arg Ser Val Gly Ile Val Arg Ser
His Lys Thr 450 455 460
Ala Ser Arg Lys Arg Tyr 465 470 231830DNAzea mays
23atgggctacc tctcctgccg cgcggactcg tccgtggcga cgtgccgctc catcacggcc
60atctcgccgc tcccactctc gcgccgctcg gggtcggggt cggggggcgg ctcgtccagg
120cccccgctgc cgccggcgca ggcggccatc gagcgcttcg actacgcgga gctggaggcg
180gccacgtccc acttcgcgga cgcggcgctg ctgggccggg gcagccacgg ggccgtctac
240aaggccgtgc tcccctcggg ccgcgccgtc gccgtcaagc gcccctcccc gcgccgcccc
300gaggtggaca acgagatccg catcctctcc tccgtccggg gcccgcggct cgtcaacctc
360ctcggcttct ctgaccccgg ccccgccccg cgcctgctcg tcgtcgagta catgcccaac
420ggcacgctct acgacctgct ccactccaac ccgcgcccgc cgggctggcc gcgccgcctg
480cgcctcgcgc tccagacggc cagggccctg cgcgcgctcc acgacgccga cccgcccgtc
540atccaccgcg acgtcaagtc cgccaacgtc ctgctcgacg ccaacctcgg cgcgcgcctc
600ggcgacttcg gcctcgccct ccgcgtgccc aaggccaccg ccggcgccaa tgccgccgcc
660gccgccgccc ccacgccgcc gcccgccggc acgctcggct acctcgaccc ggcctacgtc
720acgcccgaga gcctcagcac caagacagac gtcttcagct tcgggatcct gctgctcgag
780atcatgagcg gccgcaaggc catcgacgtc cagcactcgc cgccgtccgt cgtcgagtgg
840gccgtcccgc tcttacggaa aggcaaggtc gcctcgctgt tcgacccgcg tgtggcgccg
900ccacgagacc cggtcacccg caaggacctt gccgcgctgg ccgccagctg tgtgcgctcc
960tgcagggagc ggcgcccgtc catggccgac atcgtccagc gtctcgtgct tctcagcaaa
1020gccgtgtcgg ccaaggtgtg gaacgggctc gccgacgggc ttgccgtggt agggaaccct
1080tgtgcggttg ttgatgttca gaagaccatc tccaagcgag gtgctgccag ccgcagggct
1140gaatcagaga gggagtcgac ttcagcattg gtgtttgacg acgatgaaaa ggaggatgta
1200gatgcagagg ccttggagga agatcaggtg ccatccaaca agtcaccacc ccgaccactg
1260aagaacggga tagtgttttc cgaggcaggg gctcgggaga ggagaaatct cttggacctg
1320atggctcgga tcgatggtgt tgccggccaa agattcggca ttaccagagc aagaacagtg
1380cgtgctaatg gtgagctcat cgaaaaggat gcagtgttgc tcctaaggag gaaccagacg
1440gtaagagttg ttggatcaga ggcactgccg aagtctggaa ggatttctcg ctatgatgtg
1500aagatcaaac acaaagcagg ggaagagcaa gaggagacag ggaaagccca agacaaagta
1560gagaaaatcc aagttaatgc aagcgggatt caagaaagtt ccaaggaaat attaggcaag
1620acagataaat tgttggatgc actggagcca aaccttgaca aagaagagaa ggttcaagaa
1680aaggaagagc aacacctcga tgaagtggat aatgttcaag agaatgaagg caaagtccaa
1740tgcccggcag agaaaatcca ggaaggtgga gaagtccaat gcccccggca gagaaaatcc
1800aggaaagtcg aggattctga gacaaagtag
183024609PRTzea mays 24Met Gly Tyr Leu Ser Cys Arg Ala Asp Ser Ser Val
Ala Thr Cys Arg 1 5 10
15 Ser Ile Thr Ala Ile Ser Pro Leu Pro Leu Ser Arg Arg Ser Gly Ser
20 25 30 Gly Ser Gly
Gly Gly Ser Ser Arg Pro Pro Leu Pro Pro Ala Gln Ala 35
40 45 Ala Ile Glu Arg Phe Asp Tyr Ala
Glu Leu Glu Ala Ala Thr Ser His 50 55
60 Phe Ala Asp Ala Ala Leu Leu Gly Arg Gly Ser His Gly
Ala Val Tyr 65 70 75
80 Lys Ala Val Leu Pro Ser Gly Arg Ala Val Ala Val Lys Arg Pro Ser
85 90 95 Pro Arg Arg Pro
Glu Val Asp Asn Glu Ile Arg Ile Leu Ser Ser Val 100
105 110 Arg Gly Pro Arg Leu Val Asn Leu Leu
Gly Phe Ser Asp Pro Gly Pro 115 120
125 Ala Pro Arg Leu Leu Val Val Glu Tyr Met Pro Asn Gly Thr
Leu Tyr 130 135 140
Asp Leu Leu His Ser Asn Pro Arg Pro Pro Gly Trp Pro Arg Arg Leu 145
150 155 160 Arg Leu Ala Leu Gln
Thr Ala Arg Ala Leu Arg Ala Leu His Asp Ala 165
170 175 Asp Pro Pro Val Ile His Arg Asp Val Lys
Ser Ala Asn Val Leu Leu 180 185
190 Asp Ala Asn Leu Gly Ala Arg Leu Gly Asp Phe Gly Leu Ala Leu
Arg 195 200 205 Val
Pro Lys Ala Thr Ala Gly Ala Asn Ala Ala Ala Ala Ala Ala Pro 210
215 220 Thr Pro Pro Pro Ala Gly
Thr Leu Gly Tyr Leu Asp Pro Ala Tyr Val 225 230
235 240 Thr Pro Glu Ser Leu Ser Thr Lys Thr Asp Val
Phe Ser Phe Gly Ile 245 250
255 Leu Leu Leu Glu Ile Met Ser Gly Arg Lys Ala Ile Asp Val Gln His
260 265 270 Ser Pro
Pro Ser Val Val Glu Trp Ala Val Pro Leu Leu Arg Lys Gly 275
280 285 Lys Val Ala Ser Leu Phe Asp
Pro Arg Val Ala Pro Pro Arg Asp Pro 290 295
300 Val Thr Arg Lys Asp Leu Ala Ala Leu Ala Ala Ser
Cys Val Arg Ser 305 310 315
320 Cys Arg Glu Arg Arg Pro Ser Met Ala Asp Ile Val Gln Arg Leu Val
325 330 335 Leu Leu Ser
Lys Ala Val Ser Ala Lys Val Trp Asn Gly Leu Ala Asp 340
345 350 Gly Leu Ala Val Val Gly Asn Pro
Cys Ala Val Val Asp Val Gln Lys 355 360
365 Thr Ile Ser Lys Arg Gly Ala Ala Ser Arg Arg Ala Glu
Ser Glu Arg 370 375 380
Glu Ser Thr Ser Ala Leu Val Phe Asp Asp Asp Glu Lys Glu Asp Val 385
390 395 400 Asp Ala Glu Ala
Leu Glu Glu Asp Gln Val Pro Ser Asn Lys Ser Pro 405
410 415 Pro Arg Pro Leu Lys Asn Gly Ile Val
Phe Ser Glu Ala Gly Ala Arg 420 425
430 Glu Arg Arg Asn Leu Leu Asp Leu Met Ala Arg Ile Asp Gly
Val Ala 435 440 445
Gly Gln Arg Phe Gly Ile Thr Arg Ala Arg Thr Val Arg Ala Asn Gly 450
455 460 Glu Leu Ile Glu Lys
Asp Ala Val Leu Leu Leu Arg Arg Asn Gln Thr 465 470
475 480 Val Arg Val Val Gly Ser Glu Ala Leu Pro
Lys Ser Gly Arg Ile Ser 485 490
495 Arg Tyr Asp Val Lys Ile Lys His Lys Ala Gly Glu Glu Gln Glu
Glu 500 505 510 Thr
Gly Lys Ala Gln Asp Lys Val Glu Lys Ile Gln Val Asn Ala Ser 515
520 525 Gly Ile Gln Glu Ser Ser
Lys Glu Ile Leu Gly Lys Thr Asp Lys Leu 530 535
540 Leu Asp Ala Leu Glu Pro Asn Leu Asp Lys Glu
Glu Lys Val Gln Glu 545 550 555
560 Lys Glu Glu Gln His Leu Asp Glu Val Asp Asn Val Gln Glu Asn Glu
565 570 575 Gly Lys
Val Gln Cys Pro Ala Glu Lys Ile Gln Glu Gly Gly Glu Val 580
585 590 Gln Cys Pro Arg Gln Arg Lys
Ser Arg Lys Val Glu Asp Ser Glu Thr 595 600
605 Lys 251287DNAglycine
maxmisc_feature(747)..(747)n is a, c, g, or t 25atgccctacc tcacttgcaa
cgccgagtcc gcaatagcca catgcgaccc tcactccctc 60aagaagaaga aaaagcccaa
aagcccagcc caggcccagc ccgtgcgaca cttcgcctac 120tccgacatcc tcgacgccac
caacaacttc tctgccgaca ccttcctagg taaaggcagc 180cacggcacag tctacaaggc
cgccttccac ggcggcgctc tcgtcgccgc cgtcaaaata 240accaaaccca aaacctcaaa
cgaaatcgaa attctctccc acctcaaaaa aaaccctcgt 300cttgttaacc taattggctt
ctgcaacgac caaacccaaa ctaacaacat taacaacaac 360aaactcattg ttgtcgagta
catgccaaac ggttcgctcc acgagcttct ccactcgact 420aaaaaaccgg ttcgaccccc
aagctggacc gcgcgagtcc ggtttgcggt tcaggtcgcg 480aaagcggttc gccttttaca
ctcttccgaa ccgccagtta ttcacaggga cataaaatcg 540tccaatgtgt taatcgacga
aaagtggaac gctagactcg gtgacttcgg gctcgcggtg 600aggggacacg tggcggattc
tcgcgtgcca ccggcgggga cgttaggata cctcgacccg 660tgctatcttg cgccgggaga
tctaagttcc aagagcgatg tcttcagttt cggagttttg 720ctgctcgaga tcgcgagtgg
gaggcacgcg ctcgacgtga ggcacagtcc gccgtcggtg 780ctggactggg cggtgccgct
ggtccggcgc ggcgagttta aggagatttg tgacccgaga 840attggagcac cgccggacat
ggcggcgttc cggcggatgg cggtgctggc ggcgaggtgc 900gtgaggagca ccccggagag
aaggccgtcg atggtggagg tgttggagtg tctaacggcg 960gtgagaaaat gttttcgcgc
gccggtaatg tggaagagga ttaagaggcg cgtggagata 1020gcgcgtgggg atttgtttca
tgattgggac aggagtgagg aagttgtgag agtagttaag 1080ttaggaagta gtagtagtag
aaggaacggg aaagtatcta gtgtgtcggg tgtagagtat 1140gagggtggac acgcgaatcc
agcggtgaga tctagatcgg ttggttcggg ttcgggtttg 1200gttgggtttg ggttcaagaa
tagaaaaggg aaagtgaggc taaagagatc gaggtctatg 1260gggagtccgg tgcctctccg
gtggtga 128726428PRTglycine max
26Met Pro Tyr Leu Thr Cys Asn Ala Glu Ser Ala Ile Ala Thr Cys Asp 1
5 10 15 Pro His Ser Leu
Lys Lys Lys Lys Lys Pro Lys Ser Pro Ala Gln Ala 20
25 30 Gln Pro Val Arg His Phe Ala Tyr Ser
Asp Ile Leu Asp Ala Thr Asn 35 40
45 Asn Phe Ser Ala Asp Thr Phe Leu Gly Lys Gly Ser His Gly
Thr Val 50 55 60
Tyr Lys Ala Ala Phe His Gly Gly Ala Leu Val Ala Ala Val Lys Ile 65
70 75 80 Thr Lys Pro Lys Thr
Ser Asn Glu Ile Glu Ile Leu Ser His Leu Lys 85
90 95 Lys Asn Pro Arg Leu Val Asn Leu Ile Gly
Phe Cys Asn Asp Gln Thr 100 105
110 Gln Thr Asn Asn Ile Asn Asn Asn Lys Leu Ile Val Val Glu Tyr
Met 115 120 125 Pro
Asn Gly Ser Leu His Glu Leu Leu His Ser Thr Lys Lys Pro Val 130
135 140 Arg Pro Pro Ser Trp Thr
Ala Arg Val Arg Phe Ala Val Gln Val Ala 145 150
155 160 Lys Ala Val Arg Leu Leu His Ser Ser Glu Pro
Pro Val Ile His Arg 165 170
175 Asp Ile Lys Ser Ser Asn Val Leu Ile Asp Glu Lys Trp Asn Ala Arg
180 185 190 Leu Gly
Asp Phe Gly Leu Ala Val Arg Gly His Val Ala Asp Ser Arg 195
200 205 Val Pro Pro Ala Gly Thr Leu
Gly Tyr Leu Asp Pro Cys Tyr Leu Ala 210 215
220 Pro Gly Asp Leu Ser Ser Lys Ser Asp Val Phe Ser
Phe Gly Val Leu 225 230 235
240 Leu Leu Glu Ile Ala Ser Gly Arg His Ala Leu Asp Val Arg His Ser
245 250 255 Pro Pro Ser
Val Leu Asp Trp Ala Val Pro Leu Val Arg Arg Gly Glu 260
265 270 Phe Lys Glu Ile Cys Asp Pro Arg
Ile Gly Ala Pro Pro Asp Met Ala 275 280
285 Ala Phe Arg Arg Met Ala Val Leu Ala Ala Arg Cys Val
Arg Ser Thr 290 295 300
Pro Glu Arg Arg Pro Ser Met Val Glu Val Leu Glu Cys Leu Thr Ala 305
310 315 320 Val Arg Lys Cys
Phe Arg Ala Pro Val Met Trp Lys Arg Ile Lys Arg 325
330 335 Arg Val Glu Ile Ala Arg Gly Asp Leu
Phe His Asp Trp Asp Arg Ser 340 345
350 Glu Glu Val Val Arg Val Val Lys Leu Gly Ser Ser Ser Ser
Arg Arg 355 360 365
Asn Gly Lys Val Ser Ser Val Ser Gly Val Glu Tyr Glu Gly Gly His 370
375 380 Ala Asn Pro Ala Val
Arg Ser Arg Ser Val Gly Ser Gly Ser Gly Leu 385 390
395 400 Val Gly Phe Gly Phe Lys Asn Arg Lys Gly
Lys Val Arg Leu Lys Arg 405 410
415 Ser Arg Ser Met Gly Ser Pro Val Pro Leu Arg Trp 420
425 272040DNAoryza sativa 27atggggtacc tctcctgccg
cgcggactcg tcggtggcga cgtgccggtc catcaccgcc 60atatcgccgc tgccgctgtc
gcggcggtcg ggggtcggcg gcaggcggcg cgcgctgccg 120gcggcggcga gggaggggga
tggtggggag gcgtcgtccg ccgccgccac catcgagcgg 180ttcgcgtacg acgaactgga
ggcggcgacc tcccacttcg cggacgcggc gctgctcggg 240aggggcagcc acggggcggt
gtacaaggcg gtgctcgcct ccggccgcgc cgtcgccgtc 300aagcgcccct ccccgcgccg
ccccgaggtg gacaacgaga tccgcatcct ctcctccgtc 360cgcggcccgc gcctcgtcaa
cctcctcggc ttctccgact ccggcgccgg cgccggcgcc 420gaccagcagc agcagcagca
ccgcccgcgc ctgctcgtcg tcgagtacat gcccaacggc 480acgctctatg agctgctcca
ttccaacccg cgcccgcccg ggtggccgcg ccgcgtccgc 540ctcgcgctcc agacggcgcg
cgcgctccgc gcgctccacg acgccgatcc ccccgtcatc 600catcgcgacg tcaagtccgc
caacgtcctg ctcgacgcca acctcgacgc gcgcctcggc 660gacttcggcc tagccctccg
cgtgcccaag cggctacccg gcgacgccgc cgccaatgcc 720gccgccacgc cggcgccggc
gggcacgctc gggtacctcg acccggccta cgtcacgccg 780gagagcctca gcaccaagac
cgacgtcttc agcttcggga tcctgctgct cgagatcatg 840agcggccgca aggccatcga
cgtccagcac tcgccgccgt cggtggtgga gtgggcggtg 900cccctgctgc ggaaggggaa
ggtggcctcg ctgttcgacc cacgggtggc gccgccgcgt 960gacccggtca cccggagaga
cctagccgct ctggcggcga gctgcgtgcg gtcgtgcagg 1020gagcggcggc cgtcgatggc
cgacatagtt gatcggcttg tggttctcag caaggccgtg 1080tcgggcaaaa tgtggaacgg
actcgccgtg gttgggaacc cctgcgctgt cgtggatgtc 1140cagaagacca tcgcgaagcg
agctgctgct gctgctgcag gcgatagagc cgcgtcgcag 1200cgggagctga cttcggcatt
ggcatttgat gatgatgaga agaaagagga ggaggatgcg 1260ccgaatgcag gtgcgttaga
ggaggatgag gtgccattgg tgggtgcgaa gaaagcaccc 1320cggccattga agaatgggaa
gatgttctct gagccagggg caagggagag gagaaatctc 1380ttggagctca tggctcggat
tgatggtgtc gccggccaaa gatttggcat aactcgggca 1440agaacagtgc gtgccgctag
tgaatctatc gaaaaagatg cggcggtgtt gctcctgagg 1500agaaatcaaa ctgtgaaagt
acttggatcg gaggcccttt ctaaagctga tatcttttca 1560agtttggatg caaaaatcaa
gcatgaattg gggaaagagc agcaagagga ggcaggaaaa 1620atcaagcatg aattggtgaa
agagcagcaa gagaaggcag gaaatatcaa gcaggaattg 1680gtgaaagagc agcaagagaa
ggcaggaaat atcaagcagg aatcagggga agagcaagag 1740aaggcaggga aaaccaagca
tgatgcaggg aaagggcatg ttgagaaggc agtgggaatc 1800aatcttgaag cagggaagga
gcaggagaaa gtagagaaaa accaagagaa agaaatgaaa 1860atccaagaaa aacttgggga
aatctttgat aaagcaatga aatctgaaga aaagacaggg 1920caaaatcctg gcatagaaaa
gaaaatccaa gacacggcag agaagaaaca agagcatgat 1980gctagggtag tccaagacaa
agtggagaag atccaagacg aagccaagaa aatccaatga 204028679PRToryza sativa
28Met Gly Tyr Leu Ser Cys Arg Ala Asp Ser Ser Val Ala Thr Cys Arg 1
5 10 15 Ser Ile Thr Ala
Ile Ser Pro Leu Pro Leu Ser Arg Arg Ser Gly Val 20
25 30 Gly Gly Arg Arg Arg Ala Leu Pro Ala
Ala Ala Arg Glu Gly Asp Gly 35 40
45 Gly Glu Ala Ser Ser Ala Ala Ala Thr Ile Glu Arg Phe Ala
Tyr Asp 50 55 60
Glu Leu Glu Ala Ala Thr Ser His Phe Ala Asp Ala Ala Leu Leu Gly 65
70 75 80 Arg Gly Ser His Gly
Ala Val Tyr Lys Ala Val Leu Ala Ser Gly Arg 85
90 95 Ala Val Ala Val Lys Arg Pro Ser Pro Arg
Arg Pro Glu Val Asp Asn 100 105
110 Glu Ile Arg Ile Leu Ser Ser Val Arg Gly Pro Arg Leu Val Asn
Leu 115 120 125 Leu
Gly Phe Ser Asp Ser Gly Ala Gly Ala Gly Ala Asp Gln Gln Gln 130
135 140 Gln Gln His Arg Pro Arg
Leu Leu Val Val Glu Tyr Met Pro Asn Gly 145 150
155 160 Thr Leu Tyr Glu Leu Leu His Ser Asn Pro Arg
Pro Pro Gly Trp Pro 165 170
175 Arg Arg Val Arg Leu Ala Leu Gln Thr Ala Arg Ala Leu Arg Ala Leu
180 185 190 His Asp
Ala Asp Pro Pro Val Ile His Arg Asp Val Lys Ser Ala Asn 195
200 205 Val Leu Leu Asp Ala Asn Leu
Asp Ala Arg Leu Gly Asp Phe Gly Leu 210 215
220 Ala Leu Arg Val Pro Lys Arg Leu Pro Gly Asp Ala
Ala Ala Asn Ala 225 230 235
240 Ala Ala Thr Pro Ala Pro Ala Gly Thr Leu Gly Tyr Leu Asp Pro Ala
245 250 255 Tyr Val Thr
Pro Glu Ser Leu Ser Thr Lys Thr Asp Val Phe Ser Phe 260
265 270 Gly Ile Leu Leu Leu Glu Ile Met
Ser Gly Arg Lys Ala Ile Asp Val 275 280
285 Gln His Ser Pro Pro Ser Val Val Glu Trp Ala Val Pro
Leu Leu Arg 290 295 300
Lys Gly Lys Val Ala Ser Leu Phe Asp Pro Arg Val Ala Pro Pro Arg 305
310 315 320 Asp Pro Val Thr
Arg Arg Asp Leu Ala Ala Leu Ala Ala Ser Cys Val 325
330 335 Arg Ser Cys Arg Glu Arg Arg Pro Ser
Met Ala Asp Ile Val Asp Arg 340 345
350 Leu Val Val Leu Ser Lys Ala Val Ser Gly Lys Met Trp Asn
Gly Leu 355 360 365
Ala Val Val Gly Asn Pro Cys Ala Val Val Asp Val Gln Lys Thr Ile 370
375 380 Ala Lys Arg Ala Ala
Ala Ala Ala Ala Gly Asp Arg Ala Ala Ser Gln 385 390
395 400 Arg Glu Leu Thr Ser Ala Leu Ala Phe Asp
Asp Asp Glu Lys Lys Glu 405 410
415 Glu Glu Asp Ala Pro Asn Ala Gly Ala Leu Glu Glu Asp Glu Val
Pro 420 425 430 Leu
Val Gly Ala Lys Lys Ala Pro Arg Pro Leu Lys Asn Gly Lys Met 435
440 445 Phe Ser Glu Pro Gly Ala
Arg Glu Arg Arg Asn Leu Leu Glu Leu Met 450 455
460 Ala Arg Ile Asp Gly Val Ala Gly Gln Arg Phe
Gly Ile Thr Arg Ala 465 470 475
480 Arg Thr Val Arg Ala Ala Ser Glu Ser Ile Glu Lys Asp Ala Ala Val
485 490 495 Leu Leu
Leu Arg Arg Asn Gln Thr Val Lys Val Leu Gly Ser Glu Ala 500
505 510 Leu Ser Lys Ala Asp Ile Phe
Ser Ser Leu Asp Ala Lys Ile Lys His 515 520
525 Glu Leu Gly Lys Glu Gln Gln Glu Glu Ala Gly Lys
Ile Lys His Glu 530 535 540
Leu Val Lys Glu Gln Gln Glu Lys Ala Gly Asn Ile Lys Gln Glu Leu 545
550 555 560 Val Lys Glu
Gln Gln Glu Lys Ala Gly Asn Ile Lys Gln Glu Ser Gly 565
570 575 Glu Glu Gln Glu Lys Ala Gly Lys
Thr Lys His Asp Ala Gly Lys Gly 580 585
590 His Val Glu Lys Ala Val Gly Ile Asn Leu Glu Ala Gly
Lys Glu Gln 595 600 605
Glu Lys Val Glu Lys Asn Gln Glu Lys Glu Met Lys Ile Gln Glu Lys 610
615 620 Leu Gly Glu Ile
Phe Asp Lys Ala Met Lys Ser Glu Glu Lys Thr Gly 625 630
635 640 Gln Asn Pro Gly Ile Glu Lys Lys Ile
Gln Asp Thr Ala Glu Lys Lys 645 650
655 Gln Glu His Asp Ala Arg Val Val Gln Asp Lys Val Glu Lys
Ile Gln 660 665 670
Asp Glu Ala Lys Lys Ile Gln 675
291235DNAgossypium hirsutum 29atgggttacc tctcttgcaa tgcagagtcc accattaaag
tttgtgatcc tggcaactgg 60gattattata gaaaaaaacc caagaaaaac aagcccagaa
tccggcagtt tcgttacacc 120gatcttctta ccgccaccaa tggcttctct tccgatagct
tcctcggtaa aggtagtcac 180ggttccgtct acaaagccgt acttgacgat ggcaagttaa
tcaccgccgt taagaaaacg 240tcaaagaact gtaacagtcc tgccgacaac gagatcgaga
ttctttcccg agttgatcat 300cctcgactcg ttaatctcat cggttactgc tccgattcgc
tttgtaagaa taaattaatc 360gtcgtggaat atatgcccaa cggttcattg tacgatcttt
tacattcttc ttcttgtaaa 420ccgccgggtt ggtccagccg ggttcgattc gctttacagg
tagcaaaagc ggttcaagct 480ttacattcgg gtagcccgcc ggtgatccac agggatataa
aatcgtccaa tgttttaatt 540gatcaaaggt ggaacgctcg attgggtgat ttcgggcttg
cattgatagg acacgtggag 600gatgtacgga ttaagtgcac cccaccggcg gggacgttag
gatatctcga cccgagttat 660ttagccccga gtgacgtcag cacgaaaagt gacgtgttca
gttacggcat tttgttattg 720gagattatta gcgggaggca tgctattgat ttgaagtata
gtccgccgtc agttgttgac 780tgggcggttc cgttgataaa gaaagggaat attgttgctg
tttatgatcc aaggatttta 840cctcctaagg atcctatagt taggaagcaa ttggctgtca
ttgcagctaa atgtgtgagg 900tcttgtaggg agcgtcgccc tgcaatgaaa gaggtggtcg
gttggttaac tgggttaagc 960aaattggttc ctttacattc atggaatggt ttcagcaatc
catgtatgat ggtggaaaca 1020gtggggcgtc cggtcgattt tagaaatgcc caggagaact
tggatgcagt gcatggtacg 1080ttggctgcca aggactcgcg cagagcctat tctgatttag
gctttaggag taacttgatg 1140gaacttatgg gcatcaccag cattgatggg gaggccaaag
cttcttctag ccatagattt 1200ggtaacaaaa gctacggtaa cctttgtctc gtcct
123530411PRTgossypium hirsutum 30Met Gly Tyr Leu
Ser Cys Asn Ala Glu Ser Thr Ile Lys Val Cys Asp 1 5
10 15 Pro Gly Asn Trp Asp Tyr Tyr Arg Lys
Lys Pro Lys Lys Asn Lys Pro 20 25
30 Arg Ile Arg Gln Phe Arg Tyr Thr Asp Leu Leu Thr Ala Thr
Asn Gly 35 40 45
Phe Ser Ser Asp Ser Phe Leu Gly Lys Gly Ser His Gly Ser Val Tyr 50
55 60 Lys Ala Val Leu Asp
Asp Gly Lys Leu Ile Thr Ala Val Lys Lys Thr 65 70
75 80 Ser Lys Asn Cys Asn Ser Pro Ala Asp Asn
Glu Ile Glu Ile Leu Ser 85 90
95 Arg Val Asp His Pro Arg Leu Val Asn Leu Ile Gly Tyr Cys Ser
Asp 100 105 110 Ser
Leu Cys Lys Asn Lys Leu Ile Val Val Glu Tyr Met Pro Asn Gly 115
120 125 Ser Leu Tyr Asp Leu Leu
His Ser Ser Ser Cys Lys Pro Pro Gly Trp 130 135
140 Ser Ser Arg Val Arg Phe Ala Leu Gln Val Ala
Lys Ala Val Gln Ala 145 150 155
160 Leu His Ser Gly Ser Pro Pro Val Ile His Arg Asp Ile Lys Ser Ser
165 170 175 Asn Val
Leu Ile Asp Gln Arg Trp Asn Ala Arg Leu Gly Asp Phe Gly 180
185 190 Leu Ala Leu Ile Gly His Val
Glu Asp Val Arg Ile Lys Cys Thr Pro 195 200
205 Pro Ala Gly Thr Leu Gly Tyr Leu Asp Pro Ser Tyr
Leu Ala Pro Ser 210 215 220
Asp Val Ser Thr Lys Ser Asp Val Phe Ser Tyr Gly Ile Leu Leu Leu 225
230 235 240 Glu Ile Ile
Ser Gly Arg His Ala Ile Asp Leu Lys Tyr Ser Pro Pro 245
250 255 Ser Val Val Asp Trp Ala Val Pro
Leu Ile Lys Lys Gly Asn Ile Val 260 265
270 Ala Val Tyr Asp Pro Arg Ile Leu Pro Pro Lys Asp Pro
Ile Val Arg 275 280 285
Lys Gln Leu Ala Val Ile Ala Ala Lys Cys Val Arg Ser Cys Arg Glu 290
295 300 Arg Arg Pro Ala
Met Lys Glu Val Val Gly Trp Leu Thr Gly Leu Ser 305 310
315 320 Lys Leu Val Pro Leu His Ser Trp Asn
Gly Phe Ser Asn Pro Cys Met 325 330
335 Met Val Glu Thr Val Gly Arg Pro Val Asp Phe Arg Asn Ala
Gln Glu 340 345 350
Asn Leu Asp Ala Val His Gly Thr Leu Ala Ala Lys Asp Ser Arg Arg
355 360 365 Ala Tyr Ser Asp
Leu Gly Phe Arg Ser Asn Leu Met Glu Leu Met Gly 370
375 380 Ile Thr Ser Ile Asp Gly Glu Ala
Lys Ala Ser Ser Ser His Arg Phe 385 390
395 400 Gly Asn Lys Ser Tyr Gly Asn Leu Cys Leu Val
405 410 312025DNAArabidopsis thaliana
31atgggcatgg aagctttgag atttcttcat gttatcttct tctttgtgct aattcttcac
60tgtcattgtg gaacatctct ctctggttct tctgatgtga agcttctttt aggaaaaatc
120aagtcttcac tacaaggaaa cagtgagagc ttactgttgt cttcttggaa ctcctctgtt
180cctgtttgtc aatggagagg tgtaaaatgg gtattttcaa atgggtctcc tcttcaatgt
240agtgacctct cttcaccaca atggactaat acctctctgt tcaacgactc ttctcttcac
300cttctctctc ttcagcttcc ttctgctaat ctcactggtt cactccctag agagattggt
360gagttctcta tgcttcaaag tgtgttcctc aacatcaatt cattaagtgg gtcaatccct
420cttgagcttg gttacacttc ttctctctct gatgttgatt tgagtggtaa tgccttagct
480ggggttttgc ctccatcgat ttggaacctc tgtgataagc ttgtctcttt caagattcat
540ggtaataact tgtctggggt tttgcctgag cctgctttgc caaattcgac ttgtggtaat
600ctccaagttc ttgatttggg tggtaataag ttctcaggtg agtttcctga gtttataact
660aggtttaaag gtgtgaagtc acttgatctt tcaagtaatg tctttgaagg tcttgttcct
720gagggtttag gtgtattaga gctagaaagt ctcaatcttt ctcataataa cttcagtggg
780atgttgccag attttggtga gtcaaagttt ggagcagaat ctttcgaagg gaacagtcct
840agcctttgtg gtttgccttt gaagccttgt ctaggctcct ctaggttaag tccaggtgct
900gttgctggtc tggtgattgg tttaatgtct ggagctgttg ttgtggcctc gttgttaata
960gggtatttgc agaacaagaa aagaaagagt agtatagaga gtgaagatga tttggaagaa
1020ggtgatgaag aagatgaaat cggtgagaaa gaaggcggtg aaggaaagtt agttgtgttt
1080caaggtggtg agaatctgac gttggatgat gttttgaatg ctactgggca agttatggag
1140aagactagct atggtactgt ctataaagct aagcttagtg atggagggaa tattgcattg
1200aggctattga gagaaggtac ttgcaaggat agaagttctt gtctgcctgt tataaggcag
1260ttaggacgca ttcggcatga gaatttggtt cccttgagag ctttctatca agggaagaga
1320ggagaaaagc ttctcatcta tgactatctt cccaacataa gcttacatga tttgttgcat
1380gaaagtaaac ctcgaaagcc agctttgaat tgggctagga gacacaagat tgcacttgga
1440atagcgaggg gacttgctta tcttcatact ggacaagaag ttcctatcat ccatggaaat
1500attagatcaa agaacgtgct tgtggacgac tttttctttg caaggctaac tgagtttggg
1560cttgacaaga taatggtaca ggcagtagca gatgagattg tctcgcaggc gaaatccgac
1620gggtacaaag cacctgaact ccacaagatg aagaaatgca atccaaggag tgatgtttac
1680gcctttggga tccttctctt ggagatattg atgggtaaga aaccaggaaa gagtggaagg
1740aacggtaatg agtttgtgga cttgccttct ttggttaaag ctgcggtgtt ggaagagacg
1800acaatggagg ttttcgactt ggaggcaatg aaagggatta ggagcccaat ggaagaaggt
1860ttggttcatg cattgaagct agcgatggga tgttgtgctc ctgttacaac agttagaccc
1920agcatggaag aggttgtgaa gcagttggaa gagaacagac cgaggaatag atccgcattg
1980tacagcccaa ccgaaaccag gagcgacgcc gaaactccat tttga
202532674PRTArabidopsis thaliana 32Met Gly Met Glu Ala Leu Arg Phe Leu
His Val Ile Phe Phe Phe Val 1 5 10
15 Leu Ile Leu His Cys His Cys Gly Thr Ser Leu Ser Gly Ser
Ser Asp 20 25 30
Val Lys Leu Leu Leu Gly Lys Ile Lys Ser Ser Leu Gln Gly Asn Ser
35 40 45 Glu Ser Leu Leu
Leu Ser Ser Trp Asn Ser Ser Val Pro Val Cys Gln 50
55 60 Trp Arg Gly Val Lys Trp Val Phe
Ser Asn Gly Ser Pro Leu Gln Cys 65 70
75 80 Ser Asp Leu Ser Ser Pro Gln Trp Thr Asn Thr Ser
Leu Phe Asn Asp 85 90
95 Ser Ser Leu His Leu Leu Ser Leu Gln Leu Pro Ser Ala Asn Leu Thr
100 105 110 Gly Ser Leu
Pro Arg Glu Ile Gly Glu Phe Ser Met Leu Gln Ser Val 115
120 125 Phe Leu Asn Ile Asn Ser Leu Ser
Gly Ser Ile Pro Leu Glu Leu Gly 130 135
140 Tyr Thr Ser Ser Leu Ser Asp Val Asp Leu Ser Gly Asn
Ala Leu Ala 145 150 155
160 Gly Val Leu Pro Pro Ser Ile Trp Asn Leu Cys Asp Lys Leu Val Ser
165 170 175 Phe Lys Ile His
Gly Asn Asn Leu Ser Gly Val Leu Pro Glu Pro Ala 180
185 190 Leu Pro Asn Ser Thr Cys Gly Asn Leu
Gln Val Leu Asp Leu Gly Gly 195 200
205 Asn Lys Phe Ser Gly Glu Phe Pro Glu Phe Ile Thr Arg Phe
Lys Gly 210 215 220
Val Lys Ser Leu Asp Leu Ser Ser Asn Val Phe Glu Gly Leu Val Pro 225
230 235 240 Glu Gly Leu Gly Val
Leu Glu Leu Glu Ser Leu Asn Leu Ser His Asn 245
250 255 Asn Phe Ser Gly Met Leu Pro Asp Phe Gly
Glu Ser Lys Phe Gly Ala 260 265
270 Glu Ser Phe Glu Gly Asn Ser Pro Ser Leu Cys Gly Leu Pro Leu
Lys 275 280 285 Pro
Cys Leu Gly Ser Ser Arg Leu Ser Pro Gly Ala Val Ala Gly Leu 290
295 300 Val Ile Gly Leu Met Ser
Gly Ala Val Val Val Ala Ser Leu Leu Ile 305 310
315 320 Gly Tyr Leu Gln Asn Lys Lys Arg Lys Ser Ser
Ile Glu Ser Glu Asp 325 330
335 Asp Leu Glu Glu Gly Asp Glu Glu Asp Glu Ile Gly Glu Lys Glu Gly
340 345 350 Gly Glu
Gly Lys Leu Val Val Phe Gln Gly Gly Glu Asn Leu Thr Leu 355
360 365 Asp Asp Val Leu Asn Ala Thr
Gly Gln Val Met Glu Lys Thr Ser Tyr 370 375
380 Gly Thr Val Tyr Lys Ala Lys Leu Ser Asp Gly Gly
Asn Ile Ala Leu 385 390 395
400 Arg Leu Leu Arg Glu Gly Thr Cys Lys Asp Arg Ser Ser Cys Leu Pro
405 410 415 Val Ile Arg
Gln Leu Gly Arg Ile Arg His Glu Asn Leu Val Pro Leu 420
425 430 Arg Ala Phe Tyr Gln Gly Lys Arg
Gly Glu Lys Leu Leu Ile Tyr Asp 435 440
445 Tyr Leu Pro Asn Ile Ser Leu His Asp Leu Leu His Glu
Ser Lys Pro 450 455 460
Arg Lys Pro Ala Leu Asn Trp Ala Arg Arg His Lys Ile Ala Leu Gly 465
470 475 480 Ile Ala Arg Gly
Leu Ala Tyr Leu His Thr Gly Gln Glu Val Pro Ile 485
490 495 Ile His Gly Asn Ile Arg Ser Lys Asn
Val Leu Val Asp Asp Phe Phe 500 505
510 Phe Ala Arg Leu Thr Glu Phe Gly Leu Asp Lys Ile Met Val
Gln Ala 515 520 525
Val Ala Asp Glu Ile Val Ser Gln Ala Lys Ser Asp Gly Tyr Lys Ala 530
535 540 Pro Glu Leu His Lys
Met Lys Lys Cys Asn Pro Arg Ser Asp Val Tyr 545 550
555 560 Ala Phe Gly Ile Leu Leu Leu Glu Ile Leu
Met Gly Lys Lys Pro Gly 565 570
575 Lys Ser Gly Arg Asn Gly Asn Glu Phe Val Asp Leu Pro Ser Leu
Val 580 585 590 Lys
Ala Ala Val Leu Glu Glu Thr Thr Met Glu Val Phe Asp Leu Glu 595
600 605 Ala Met Lys Gly Ile Arg
Ser Pro Met Glu Glu Gly Leu Val His Ala 610 615
620 Leu Lys Leu Ala Met Gly Cys Cys Ala Pro Val
Thr Thr Val Arg Pro 625 630 635
640 Ser Met Glu Glu Val Val Lys Gln Leu Glu Glu Asn Arg Pro Arg Asn
645 650 655 Arg Ser
Ala Leu Tyr Ser Pro Thr Glu Thr Arg Ser Asp Ala Glu Thr 660
665 670 Pro Phe 332037DNAzea mays
33atgcatcctt gcatgctctt cctcctctgc ctggccgcgc tgccgctggc ctcgcattcc
60tcctccaacc ccgacgtcgc gctgctcctc gccaaggtga agccggcgct gcagggcgag
120cgcgccaacg cgcagctcgc cacctggaac gcctccacgc cgctctgcct ctggcgcggc
180ctccgctggg cgacgcccga cggccggccc ctccgctgcg acgccgccgc cacgcgcgcc
240aacctgtcgc tcgcctccga ccccgccctc ctcctcctct ccgtccgcct ccccgcgtcc
300gccctcgccg gccgcctccc gccggacctc ggcgccttct ccgcgctcga ctccgtctac
360ctcgccgcca actcgctctc gggccccgtc ccgctcgagc tcggcaacgc gcccgcgctc
420tccgcgctcg acctcgccgg caaccgcctc tcgggggacc tgcccgcctc catctggaac
480ctctgcgacc gcgccaccga gctccgcctc cacggcaacg cgctcaccgg ggccgtgccc
540gagccggccg gccccaacac cacctgcgac cgcctccgcg tcctcgacct cggcgccaac
600cgcttctccg gcgccttccc cgccttcgtc accgcgttcc gtggcctcca gcgcctcgac
660ctgggcgcca accgcctgga gggccccatc ccggaggccc tcgctgggat ggcggcgacc
720cagcagctcc aggcgctcaa cgtctcctac aacaacttct ccggccagct gcccccgtcc
780ttcgcggcct cccgcttcac ggcggactcg ttcgtaggca acgaaccagc gctgtgcggc
840ccgccgctgc gccagtgcgt gacagcctcg ggcctcagct cccgcggcgt cgccgggatg
900gtcatcggga tcatggccgg cgccgtcgtg ctcgcgtccg tgtccatcgg ctgggcgcag
960gggaggtgga ggcggagcgg caggatcccg gagcaggacg agatgctgga gtcggccgac
1020gacgcccagg acgcgtcgtc agagggcagg ctcgtggtct tcgagggcgg cgagcacctc
1080acgctggagg aggtgctcaa cgcgaccggc caggtggtgg acaaggccag ctactgcacg
1140gtgtacaagg cgaagctggc gagcggcggc agcagcatcg agctgcgcct gctgcgggaa
1200ggcagctgca aggacgccgc gtcgtgcgcg cccgttgtgc ggcgcatcgg ccgcgcgcgc
1260cacgagaacc tggtcccgct tagggccttc taccagggga ggcgcggcga gaagctgctg
1320gtgtacgact acttcccgcg cagccggacg ctgcaggagc tgctgcacgg tggcagcgag
1380cccgcggcgg ggcggccggc gctcacctgg gggcggcggc acaagatcgc gctgggcgcg
1440gcgcgcgcgc tggcgtatct gcacgcgggc cagggcgagg cgcacgggaa cgtgcgctcg
1500tccatcgtgg tggtggacga cctcttcgtg ccgcgcctgg cagagtacgc ggtggaccgg
1560ctgctggtgc cggcggcggc ggaggcggtg ctggcggcgg ccaaggcgga cgggtacaag
1620gcgccggagc tgcactccat gaagaagtgc agcgcgcgca cggacgtgta cgcgttcggg
1680atcctgctgc tggagctgct catggggagg aagccgtcgg cctctgcagg tggagctgca
1740agggcgatgg acctgccgtc ggtggtgaag gtggcggtgc tggaggagac ggcgctggag
1800gaggtgctgg acgcggaggt ggtcaaggga ctgcgggtga gtccggcgga ggaggggctg
1860gtgcaggcgc tgaagctggc gatgggctgc tgcgcgccgg tgccagcggc gaggccgagc
1920atggcggagg tggtgcggca gctggaggag agccggccca agaacgtcca cccgcggtct
1980gcgctgtaca gccctacgga gagcaggagc gacgccggca cgccgaccac cgcctag
203734678PRTzea mays 34Met His Pro Cys Met Leu Phe Leu Leu Cys Leu Ala
Ala Leu Pro Leu 1 5 10
15 Ala Ser His Ser Ser Ser Asn Pro Asp Val Ala Leu Leu Leu Ala Lys
20 25 30 Val Lys Pro
Ala Leu Gln Gly Glu Arg Ala Asn Ala Gln Leu Ala Thr 35
40 45 Trp Asn Ala Ser Thr Pro Leu Cys
Leu Trp Arg Gly Leu Arg Trp Ala 50 55
60 Thr Pro Asp Gly Arg Pro Leu Arg Cys Asp Ala Ala Ala
Thr Arg Ala 65 70 75
80 Asn Leu Ser Leu Ala Ser Asp Pro Ala Leu Leu Leu Leu Ser Val Arg
85 90 95 Leu Pro Ala Ser
Ala Leu Ala Gly Arg Leu Pro Pro Asp Leu Gly Ala 100
105 110 Phe Ser Ala Leu Asp Ser Val Tyr Leu
Ala Ala Asn Ser Leu Ser Gly 115 120
125 Pro Val Pro Leu Glu Leu Gly Asn Ala Pro Ala Leu Ser Ala
Leu Asp 130 135 140
Leu Ala Gly Asn Arg Leu Ser Gly Asp Leu Pro Ala Ser Ile Trp Asn 145
150 155 160 Leu Cys Asp Arg Ala
Thr Glu Leu Arg Leu His Gly Asn Ala Leu Thr 165
170 175 Gly Ala Val Pro Glu Pro Ala Gly Pro Asn
Thr Thr Cys Asp Arg Leu 180 185
190 Arg Val Leu Asp Leu Gly Ala Asn Arg Phe Ser Gly Ala Phe Pro
Ala 195 200 205 Phe
Val Thr Ala Phe Arg Gly Leu Gln Arg Leu Asp Leu Gly Ala Asn 210
215 220 Arg Leu Glu Gly Pro Ile
Pro Glu Ala Leu Ala Gly Met Ala Ala Thr 225 230
235 240 Gln Gln Leu Gln Ala Leu Asn Val Ser Tyr Asn
Asn Phe Ser Gly Gln 245 250
255 Leu Pro Pro Ser Phe Ala Ala Ser Arg Phe Thr Ala Asp Ser Phe Val
260 265 270 Gly Asn
Glu Pro Ala Leu Cys Gly Pro Pro Leu Arg Gln Cys Val Thr 275
280 285 Ala Ser Gly Leu Ser Ser Arg
Gly Val Ala Gly Met Val Ile Gly Ile 290 295
300 Met Ala Gly Ala Val Val Leu Ala Ser Val Ser Ile
Gly Trp Ala Gln 305 310 315
320 Gly Arg Trp Arg Arg Ser Gly Arg Ile Pro Glu Gln Asp Glu Met Leu
325 330 335 Glu Ser Ala
Asp Asp Ala Gln Asp Ala Ser Ser Glu Gly Arg Leu Val 340
345 350 Val Phe Glu Gly Gly Glu His Leu
Thr Leu Glu Glu Val Leu Asn Ala 355 360
365 Thr Gly Gln Val Val Asp Lys Ala Ser Tyr Cys Thr Val
Tyr Lys Ala 370 375 380
Lys Leu Ala Ser Gly Gly Ser Ser Ile Glu Leu Arg Leu Leu Arg Glu 385
390 395 400 Gly Ser Cys Lys
Asp Ala Ala Ser Cys Ala Pro Val Val Arg Arg Ile 405
410 415 Gly Arg Ala Arg His Glu Asn Leu Val
Pro Leu Arg Ala Phe Tyr Gln 420 425
430 Gly Arg Arg Gly Glu Lys Leu Leu Val Tyr Asp Tyr Phe Pro
Arg Ser 435 440 445
Arg Thr Leu Gln Glu Leu Leu His Gly Gly Ser Glu Pro Ala Ala Gly 450
455 460 Arg Pro Ala Leu Thr
Trp Gly Arg Arg His Lys Ile Ala Leu Gly Ala 465 470
475 480 Ala Arg Ala Leu Ala Tyr Leu His Ala Gly
Gln Gly Glu Ala His Gly 485 490
495 Asn Val Arg Ser Ser Ile Val Val Val Asp Asp Leu Phe Val Pro
Arg 500 505 510 Leu
Ala Glu Tyr Ala Val Asp Arg Leu Leu Val Pro Ala Ala Ala Glu 515
520 525 Ala Val Leu Ala Ala Ala
Lys Ala Asp Gly Tyr Lys Ala Pro Glu Leu 530 535
540 His Ser Met Lys Lys Cys Ser Ala Arg Thr Asp
Val Tyr Ala Phe Gly 545 550 555
560 Ile Leu Leu Leu Glu Leu Leu Met Gly Arg Lys Pro Ser Ala Ser Ala
565 570 575 Gly Gly
Ala Ala Arg Ala Met Asp Leu Pro Ser Val Val Lys Val Ala 580
585 590 Val Leu Glu Glu Thr Ala Leu
Glu Glu Val Leu Asp Ala Glu Val Val 595 600
605 Lys Gly Leu Arg Val Ser Pro Ala Glu Glu Gly Leu
Val Gln Ala Leu 610 615 620
Lys Leu Ala Met Gly Cys Cys Ala Pro Val Pro Ala Ala Arg Pro Ser 625
630 635 640 Met Ala Glu
Val Val Arg Gln Leu Glu Glu Ser Arg Pro Lys Asn Val 645
650 655 His Pro Arg Ser Ala Leu Tyr Ser
Pro Thr Glu Ser Arg Ser Asp Ala 660 665
670 Gly Thr Pro Thr Thr Ala 675
352100DNAglycine max 35atggcgtttc tgaacccttt ctctctccac attcccatgt
ccttgttgtt cttcttcctc 60ttcttttttt gcttctgcaa agccagtagt actaccacct
ccaccacaaa gtcactgtcc 120cccccttctt caccctccac tacttcctcc tctgacgttg
agcttctctt gggaaagatc 180aaagcttcac tgcaaggtag taactctgac aaccttgttt
tgtcttcatg gaactcctcc 240accccacttt gtcagtggaa aggcctcata tgggtcttct
ccaatggcac tcctctctca 300tgcactgact tgtcctctcc tcaatggacc aatctcacac
tcctcaaaga cccttctctt 360cacttgtttt ccctccggct cccttctgca aacctctctg
gttccctccc tagagagctt 420ggagggttcc ctatgctcca aagtctctac cttaacatta
actcattgga gggtaccatc 480cctcttgagc ttggttatag ctcctctctc tctgagattg
atttgggtga caatatgcta 540ggtggggttc ttccaccttc tatttggaac ttgtgtgaga
ggcttgtttc ccttaggctc 600cacggtaatt ctttatctgg gttagtttct gagcctgcat
tgcctaactc ttcttgcaag 660aatctgcagg tgcttgattt gggtggcaac aagttctctg
ggagtttccc tgagttcatc 720acaaagtttg gtggcctaaa gcagcttgac ttggggaata
acatgtttat gggtgcaatt 780cctcaaggcc tagctgggct tagtcttgaa aaattgaatc
tttcacacaa taactttagt 840ggggttttgc ctttgtttgg aggagaatcc aagtttggtg
tggatgcttt tgaggggaat 900agccctagcc tgtgtggacc acctctggga agctgtgcta
ggacctctac actgagttct 960ggtgctgttg ctggcattgt tattagtctg atgacaggag
ctgtggtttt ggcttctttg 1020ctgatagggt atatgcagaa caagaagaag aaggggagtg
gggagagtga ggatgagttg 1080aatgatgaag aggaagatga tgaagagaat ggtggtaatg
ctattggtgg agctggtgag 1140gggaagctca tgttatttgc tggaggtgag aatttgacat
tggatgatgt gttgaatgca 1200actgggcagg ttttggagaa gacttgttat gggacggctt
ataaggctaa gcttgctgat 1260ggaggcacca ttgctttgag gctgttgaga gaaggtagct
gcaaagacaa ggcttcttgc 1320ttgtctgtta taaagcaatt ggggaaaatt cgccacgaga
atttgattcc tttgagagct 1380ttctatcagg ggaagagagg ggagaagctc cttatttatg
actacctgcc tctcagaacc 1440cttcatgatc ttttacatgg agctaaagct ggaaaaccag
tgttgaactg ggctaggcga 1500cacaagattg cgctgggcat agcgagaggt ctagcttatc
ttcacacagg acttgaagtt 1560cctgtcaccc atgcaaacgt aaggtccaag aatgtgcttg
tggatgactt ctttacagcc 1620aggctcaccg attttggtct tgacaagctg atgattcctt
ccatagccga cgaaatggta 1680gcgcttgcta agacggacgg ctacaaggct cctgagcttc
aaagaatgaa gaaatgcaac 1740tccaggactg atgtttatgc attcggcata ctgctgcttg
aaatcttgat tgggaagaag 1800cctgggaaga atggaagaaa tggcgagtat gtggacttgc
cttcgatggt gaaagtggcg 1860gttttggagg agacgacgat ggaagtgttt gatgtggagc
ttttgaaagg gataagaagc 1920cctatggaag atgggttggt gcaggcgctg aagctggcaa
tggggtgctg tgcaccagtg 1980gcatctgtta ggccaagcat ggatgaagtt gtgaggcagt
tggaggagaa tagaccaagg 2040aacaggtctg cattatacag ccctacagaa acaagaagtg
gaagtgttac cccattttga 210036699PRTglycine max 36Met Ala Phe Leu Asn Pro
Phe Ser Leu His Ile Pro Met Ser Leu Leu 1 5
10 15 Phe Phe Phe Leu Phe Phe Phe Cys Phe Cys Lys
Ala Ser Ser Thr Thr 20 25
30 Thr Ser Thr Thr Lys Ser Leu Ser Pro Pro Ser Ser Pro Ser Thr
Thr 35 40 45 Ser
Ser Ser Asp Val Glu Leu Leu Leu Gly Lys Ile Lys Ala Ser Leu 50
55 60 Gln Gly Ser Asn Ser Asp
Asn Leu Val Leu Ser Ser Trp Asn Ser Ser 65 70
75 80 Thr Pro Leu Cys Gln Trp Lys Gly Leu Ile Trp
Val Phe Ser Asn Gly 85 90
95 Thr Pro Leu Ser Cys Thr Asp Leu Ser Ser Pro Gln Trp Thr Asn Leu
100 105 110 Thr Leu
Leu Lys Asp Pro Ser Leu His Leu Phe Ser Leu Arg Leu Pro 115
120 125 Ser Ala Asn Leu Ser Gly Ser
Leu Pro Arg Glu Leu Gly Gly Phe Pro 130 135
140 Met Leu Gln Ser Leu Tyr Leu Asn Ile Asn Ser Leu
Glu Gly Thr Ile 145 150 155
160 Pro Leu Glu Leu Gly Tyr Ser Ser Ser Leu Ser Glu Ile Asp Leu Gly
165 170 175 Asp Asn Met
Leu Gly Gly Val Leu Pro Pro Ser Ile Trp Asn Leu Cys 180
185 190 Glu Arg Leu Val Ser Leu Arg Leu
His Gly Asn Ser Leu Ser Gly Leu 195 200
205 Val Ser Glu Pro Ala Leu Pro Asn Ser Ser Cys Lys Asn
Leu Gln Val 210 215 220
Leu Asp Leu Gly Gly Asn Lys Phe Ser Gly Ser Phe Pro Glu Phe Ile 225
230 235 240 Thr Lys Phe Gly
Gly Leu Lys Gln Leu Asp Leu Gly Asn Asn Met Phe 245
250 255 Met Gly Ala Ile Pro Gln Gly Leu Ala
Gly Leu Ser Leu Glu Lys Leu 260 265
270 Asn Leu Ser His Asn Asn Phe Ser Gly Val Leu Pro Leu Phe
Gly Gly 275 280 285
Glu Ser Lys Phe Gly Val Asp Ala Phe Glu Gly Asn Ser Pro Ser Leu 290
295 300 Cys Gly Pro Pro Leu
Gly Ser Cys Ala Arg Thr Ser Thr Leu Ser Ser 305 310
315 320 Gly Ala Val Ala Gly Ile Val Ile Ser Leu
Met Thr Gly Ala Val Val 325 330
335 Leu Ala Ser Leu Leu Ile Gly Tyr Met Gln Asn Lys Lys Lys Lys
Gly 340 345 350 Ser
Gly Glu Ser Glu Asp Glu Leu Asn Asp Glu Glu Glu Asp Asp Glu 355
360 365 Glu Asn Gly Gly Asn Ala
Ile Gly Gly Ala Gly Glu Gly Lys Leu Met 370 375
380 Leu Phe Ala Gly Gly Glu Asn Leu Thr Leu Asp
Asp Val Leu Asn Ala 385 390 395
400 Thr Gly Gln Val Leu Glu Lys Thr Cys Tyr Gly Thr Ala Tyr Lys Ala
405 410 415 Lys Leu
Ala Asp Gly Gly Thr Ile Ala Leu Arg Leu Leu Arg Glu Gly 420
425 430 Ser Cys Lys Asp Lys Ala Ser
Cys Leu Ser Val Ile Lys Gln Leu Gly 435 440
445 Lys Ile Arg His Glu Asn Leu Ile Pro Leu Arg Ala
Phe Tyr Gln Gly 450 455 460
Lys Arg Gly Glu Lys Leu Leu Ile Tyr Asp Tyr Leu Pro Leu Arg Thr 465
470 475 480 Leu His Asp
Leu Leu His Gly Ala Lys Ala Gly Lys Pro Val Leu Asn 485
490 495 Trp Ala Arg Arg His Lys Ile Ala
Leu Gly Ile Ala Arg Gly Leu Ala 500 505
510 Tyr Leu His Thr Gly Leu Glu Val Pro Val Thr His Ala
Asn Val Arg 515 520 525
Ser Lys Asn Val Leu Val Asp Asp Phe Phe Thr Ala Arg Leu Thr Asp 530
535 540 Phe Gly Leu Asp
Lys Leu Met Ile Pro Ser Ile Ala Asp Glu Met Val 545 550
555 560 Ala Leu Ala Lys Thr Asp Gly Tyr Lys
Ala Pro Glu Leu Gln Arg Met 565 570
575 Lys Lys Cys Asn Ser Arg Thr Asp Val Tyr Ala Phe Gly Ile
Leu Leu 580 585 590
Leu Glu Ile Leu Ile Gly Lys Lys Pro Gly Lys Asn Gly Arg Asn Gly
595 600 605 Glu Tyr Val Asp
Leu Pro Ser Met Val Lys Val Ala Val Leu Glu Glu 610
615 620 Thr Thr Met Glu Val Phe Asp Val
Glu Leu Leu Lys Gly Ile Arg Ser 625 630
635 640 Pro Met Glu Asp Gly Leu Val Gln Ala Leu Lys Leu
Ala Met Gly Cys 645 650
655 Cys Ala Pro Val Ala Ser Val Arg Pro Ser Met Asp Glu Val Val Arg
660 665 670 Gln Leu Glu
Glu Asn Arg Pro Arg Asn Arg Ser Ala Leu Tyr Ser Pro 675
680 685 Thr Glu Thr Arg Ser Gly Ser Val
Thr Pro Phe 690 695 371989DNAoryza
sativa 37atgagatctg tgatgatgtg ctgcctcctc ctcctcctgg tctctgccgc
tgccggggcc 60gagggcaagt cggaggtggc gctcctcctg gagcgcgtga agccggcgct
gcagggggag 120ggcgaagtag gagggaacgc gcagctggcc acctggaccg cctccacccc
gctctgccag 180tggcgcggcc tccgctggtc caccgccgcc accctccccc gcgagctccc
ctgcggcaac 240ctctctgcag gcctcgccca ccacccggtc ccggacgacc tcctcctcct
cctctccatc 300cgcctcccgg cctccgccct cgccggccac ctccctcccg aactcgccgc
tttctccgcc 360ctcgcctcca tcttcctcgc ccacaactcc ctctccgggc ccatccccct
cgccctcggc 420aacgcccccg ccctctccct cctcgacctc gcctccaacc gcctctccgg
ctccctcccg 480ctctccatct ggaacctctg cagcggcaac gcccgtctct ccctcctccg
cctccacggc 540aacgccctcc acggcccaat ccccgacccc gccgccctcg cccccaacac
cacctgcgac 600gccctcagcc tcctcgacct ctccgccaac cgcctctccg gccccttccc
ctcctcccta 660gtcaccaccg ccttccccgc cctccgctcc ctcgacctct ccgacaaccg
cctccacggt 720cccatcccgc acggcctcgc ccccatccac tccctcaacc tctcctacaa
caacttctcc 780ggccaacttc cccccgacct cgcctctctg ccgcccgacg ccttcctcgc
caacagcccc 840gcgctctgcg gcccgccgct gccccaccac tgcctcccca gcaaccccct
cacctcctcc 900gccgtcgccg ccatcgtcat tgccctcatg gccgccgccg tcgtcctggc
ttccctctcc 960atcggctggg cgcagggccg ttggaggcga gcgcctttgc cgccggagga
agggacactc 1020acggaggacg gcgaggggaa gctggtggtg ttccagggcg gggagcacct
cacgctggag 1080gaggtgctca acgccacggg gcaggtggtc aacaaggcca gctactgcac
cgtctacaag 1140gccaagctgg cggagggcgg cggcagcatc gagctgcgcc tcctccgcga
gggctgctgc 1200aaggacgccg agtcgtgcgc gccggcggtg cgccgcatcg gccgcgcgcg
ccacgacaac 1260ctggttccgc tgcgcgcctt ctaccagggg cgccgcggag agaagctgct
ggtgtacgac 1320tacttccccg gcaaccggac gctccacgag ctcctccacg gccacgggga
gcagagccag 1380gggatgaggc cggcgttgac gtgggcgcgg cggcacaaga tcgcgctggg
cgtggcgcgc 1440gcgctggcgt acgtgcacgc ggggcacggc gaggcgcacg gcagcgtgcg
ctcgtccaac 1500gtgctggtgg acgagtggtt cgtggcgagg gtggcggagt acgcagtgca
ccggctgctg 1560gtggcggcgg cggtggggaa ggcggacggg tacagggcgc cggagctgca
gtcgaggggg 1620aggtgcagcc cgcggacgga cgtgtacgcg ttcgggatat tgctgctgga
gctgctgatg 1680gggcggaagg cgtcgggaga gctgccggcg gtggtgaagg cggcggtgct
ggaggaggtg 1740acgatgatgg aggtgttcga cgcggaggtg gcgcgcgggg tgcgtagccc
cgcggaggag 1800gggctgcttc aggcgctgaa gctggcgatg gggtgctgcg cgccggtggc
ttcggcaagg 1860cccaccatgg cggaggtggt gcggcagctg gaggaggtcc ggccccggaa
cagcagccgg 1920ccgtcggcga tctacagccc cgccgagccc aggagcgacg ccggcacgcc
caccgccgcc 1980gccgtctaa
198938662PRToryza sativa 38Met Arg Ser Val Met Met Cys Cys Leu
Leu Leu Leu Leu Val Ser Ala 1 5 10
15 Ala Ala Gly Ala Glu Gly Lys Ser Glu Val Ala Leu Leu Leu
Glu Arg 20 25 30
Val Lys Pro Ala Leu Gln Gly Glu Gly Glu Val Gly Gly Asn Ala Gln
35 40 45 Leu Ala Thr Trp
Thr Ala Ser Thr Pro Leu Cys Gln Trp Arg Gly Leu 50
55 60 Arg Trp Ser Thr Ala Ala Thr Leu
Pro Arg Glu Leu Pro Cys Gly Asn 65 70
75 80 Leu Ser Ala Gly Leu Ala His His Pro Val Pro Asp
Asp Leu Leu Leu 85 90
95 Leu Leu Ser Ile Arg Leu Pro Ala Ser Ala Leu Ala Gly His Leu Pro
100 105 110 Pro Glu Leu
Ala Ala Phe Ser Ala Leu Ala Ser Ile Phe Leu Ala His 115
120 125 Asn Ser Leu Ser Gly Pro Ile Pro
Leu Ala Leu Gly Asn Ala Pro Ala 130 135
140 Leu Ser Leu Leu Asp Leu Ala Ser Asn Arg Leu Ser Gly
Ser Leu Pro 145 150 155
160 Leu Ser Ile Trp Asn Leu Cys Ser Gly Asn Ala Arg Leu Ser Leu Leu
165 170 175 Arg Leu His Gly
Asn Ala Leu His Gly Pro Ile Pro Asp Pro Ala Ala 180
185 190 Leu Ala Pro Asn Thr Thr Cys Asp Ala
Leu Ser Leu Leu Asp Leu Ser 195 200
205 Ala Asn Arg Leu Ser Gly Pro Phe Pro Ser Ser Leu Val Thr
Thr Ala 210 215 220
Phe Pro Ala Leu Arg Ser Leu Asp Leu Ser Asp Asn Arg Leu His Gly 225
230 235 240 Pro Ile Pro His Gly
Leu Ala Pro Ile His Ser Leu Asn Leu Ser Tyr 245
250 255 Asn Asn Phe Ser Gly Gln Leu Pro Pro Asp
Leu Ala Ser Leu Pro Pro 260 265
270 Asp Ala Phe Leu Ala Asn Ser Pro Ala Leu Cys Gly Pro Pro Leu
Pro 275 280 285 His
His Cys Leu Pro Ser Asn Pro Leu Thr Ser Ser Ala Val Ala Ala 290
295 300 Ile Val Ile Ala Leu Met
Ala Ala Ala Val Val Leu Ala Ser Leu Ser 305 310
315 320 Ile Gly Trp Ala Gln Gly Arg Trp Arg Arg Ala
Pro Leu Pro Pro Glu 325 330
335 Glu Gly Thr Leu Thr Glu Asp Gly Glu Gly Lys Leu Val Val Phe Gln
340 345 350 Gly Gly
Glu His Leu Thr Leu Glu Glu Val Leu Asn Ala Thr Gly Gln 355
360 365 Val Val Asn Lys Ala Ser Tyr
Cys Thr Val Tyr Lys Ala Lys Leu Ala 370 375
380 Glu Gly Gly Gly Ser Ile Glu Leu Arg Leu Leu Arg
Glu Gly Cys Cys 385 390 395
400 Lys Asp Ala Glu Ser Cys Ala Pro Ala Val Arg Arg Ile Gly Arg Ala
405 410 415 Arg His Asp
Asn Leu Val Pro Leu Arg Ala Phe Tyr Gln Gly Arg Arg 420
425 430 Gly Glu Lys Leu Leu Val Tyr Asp
Tyr Phe Pro Gly Asn Arg Thr Leu 435 440
445 His Glu Leu Leu His Gly His Gly Glu Gln Ser Gln Gly
Met Arg Pro 450 455 460
Ala Leu Thr Trp Ala Arg Arg His Lys Ile Ala Leu Gly Val Ala Arg 465
470 475 480 Ala Leu Ala Tyr
Val His Ala Gly His Gly Glu Ala His Gly Ser Val 485
490 495 Arg Ser Ser Asn Val Leu Val Asp Glu
Trp Phe Val Ala Arg Val Ala 500 505
510 Glu Tyr Ala Val His Arg Leu Leu Val Ala Ala Ala Val Gly
Lys Ala 515 520 525
Asp Gly Tyr Arg Ala Pro Glu Leu Gln Ser Arg Gly Arg Cys Ser Pro 530
535 540 Arg Thr Asp Val Tyr
Ala Phe Gly Ile Leu Leu Leu Glu Leu Leu Met 545 550
555 560 Gly Arg Lys Ala Ser Gly Glu Leu Pro Ala
Val Val Lys Ala Ala Val 565 570
575 Leu Glu Glu Val Thr Met Met Glu Val Phe Asp Ala Glu Val Ala
Arg 580 585 590 Gly
Val Arg Ser Pro Ala Glu Glu Gly Leu Leu Gln Ala Leu Lys Leu 595
600 605 Ala Met Gly Cys Cys Ala
Pro Val Ala Ser Ala Arg Pro Thr Met Ala 610 615
620 Glu Val Val Arg Gln Leu Glu Glu Val Arg Pro
Arg Asn Ser Ser Arg 625 630 635
640 Pro Ser Ala Ile Tyr Ser Pro Ala Glu Pro Arg Ser Asp Ala Gly Thr
645 650 655 Pro Thr
Ala Ala Ala Val 660 391254DNAGossypium hirsutum
39tcccttgatt taggtaacaa caagttttta ggggatttcc cagagtttgt aactaggttt
60caagctctta gagagcttga tctttcgagt aacatgcttt caggtcaaat tccacagagt
120ttggccactt taaacgtgga aaaattaaac ctttcccaca ataacttcac tggaatgttg
180cctgtttttg gtgaaagaaa gtttggcccg gaggctttcg aagggaacaa tccagggcta
240tgtgggttgc ctttgaatag ttgtagtggc aggtcacagc tgagtccagg tgcaattgct
300ggcattgtga ttggtctaat gactggagtg gtagttttgg catcattgtt cgttggctat
360atgcaaaaca ggaagaggag cagcaatgga gatagtgagg aggaactgga agaaggagag
420ggggatgaaa acggggtcgg gggagttgtc agcgagagca agcttatttt gtttcaaggc
480ggggagcatt tgacattaga ggatgtactg aatgcaactg gtcaagtcat ggagaagaca
540aattatggga ctgtttataa ggcaaagctt gctgatggtg gaaatatagc attgaggttg
600ttgagggaag gcagttgtaa ggacgggagt tcatgtctgc ctgtcataaa gcagctgggg
660aaggttagac atgagaattt ggttccactg agagcattct atcaggggaa aagaggggaa
720aagcttctaa tttatgacta tcttccaaat agaagcctac atgacttttt acatggtatg
780caagcaggaa agccagttct aaattgggct cgacggcaca aaatcgcatt ggggatagcc
840aaaggattag cacatcttca tacaggtctc gagatgccga tcacccatgg gaatgttagg
900tccaaaaatg tgcttgtaga tgacttcttt gtagccaggc tcaccgaata tggactcgac
960aagctaatga tcccggctgt ggctgatgaa atggttgccc tcgcaaagac cgaatgttac
1020aaggcaccgg aacttcaaag catgaagaaa tgcaacacca gaactgacgt ttatgcattt
1080gggatattgt tattagagat tttgataggc aagaagcctg agaaaaatgc aagacgcaac
1140gatgttgggg atttgccttc gattgtgaaa gcagcagttt tggaagagac aacaatggag
1200gttttcgacg tagaagtgtt gaaggtattc gaagtccgat ggaagacggg atag
125440417PRTGossypium hirsutum 40Ser Leu Asp Leu Gly Asn Asn Lys Phe Leu
Gly Asp Phe Pro Glu Phe 1 5 10
15 Val Thr Arg Phe Gln Ala Leu Arg Glu Leu Asp Leu Ser Ser Asn
Met 20 25 30 Leu
Ser Gly Gln Ile Pro Gln Ser Leu Ala Thr Leu Asn Val Glu Lys 35
40 45 Leu Asn Leu Ser His Asn
Asn Phe Thr Gly Met Leu Pro Val Phe Gly 50 55
60 Glu Arg Lys Phe Gly Pro Glu Ala Phe Glu Gly
Asn Asn Pro Gly Leu 65 70 75
80 Cys Gly Leu Pro Leu Asn Ser Cys Ser Gly Arg Ser Gln Leu Ser Pro
85 90 95 Gly Ala
Ile Ala Gly Ile Val Ile Gly Leu Met Thr Gly Val Val Val 100
105 110 Leu Ala Ser Leu Phe Val Gly
Tyr Met Gln Asn Arg Lys Arg Ser Ser 115 120
125 Asn Gly Asp Ser Glu Glu Glu Leu Glu Glu Gly Glu
Gly Asp Glu Asn 130 135 140
Gly Val Gly Gly Val Val Ser Glu Ser Lys Leu Ile Leu Phe Gln Gly 145
150 155 160 Gly Glu His
Leu Thr Leu Glu Asp Val Leu Asn Ala Thr Gly Gln Val 165
170 175 Met Glu Lys Thr Asn Tyr Gly Thr
Val Tyr Lys Ala Lys Leu Ala Asp 180 185
190 Gly Gly Asn Ile Ala Leu Arg Leu Leu Arg Glu Gly Ser
Cys Lys Asp 195 200 205
Gly Ser Ser Cys Leu Pro Val Ile Lys Gln Leu Gly Lys Val Arg His 210
215 220 Glu Asn Leu Val
Pro Leu Arg Ala Phe Tyr Gln Gly Lys Arg Gly Glu 225 230
235 240 Lys Leu Leu Ile Tyr Asp Tyr Leu Pro
Asn Arg Ser Leu His Asp Phe 245 250
255 Leu His Gly Met Gln Ala Gly Lys Pro Val Leu Asn Trp Ala
Arg Arg 260 265 270
His Lys Ile Ala Leu Gly Ile Ala Lys Gly Leu Ala His Leu His Thr
275 280 285 Gly Leu Glu Met
Pro Ile Thr His Gly Asn Val Arg Ser Lys Asn Val 290
295 300 Leu Val Asp Asp Phe Phe Val Ala
Arg Leu Thr Glu Tyr Gly Leu Asp 305 310
315 320 Lys Leu Met Ile Pro Ala Val Ala Asp Glu Met Val
Ala Leu Ala Lys 325 330
335 Thr Glu Cys Tyr Lys Ala Pro Glu Leu Gln Ser Met Lys Lys Cys Asn
340 345 350 Thr Arg Thr
Asp Val Tyr Ala Phe Gly Ile Leu Leu Leu Glu Ile Leu 355
360 365 Ile Gly Lys Lys Pro Glu Lys Asn
Ala Arg Arg Asn Asp Val Gly Asp 370 375
380 Leu Pro Ser Ile Val Lys Ala Ala Val Leu Glu Glu Thr
Thr Met Glu 385 390 395
400 Val Phe Asp Val Glu Val Leu Lys Val Phe Glu Val Arg Trp Lys Thr
405 410 415 Gly
411263DNAarabidopsis thaliana 41atgaaatgct tcttattccc tcttggggac
aagaaagatg aacagagaag ccctaaaccg 60gtttcaccaa cgtctaactt cagtgacgta
aacaaaagcg gttcagattt cagtccccgg 120gatgtttctg gaacgagcac agtatcatcc
actggtagga actcgaacac tagcatgtca 180gctagagaaa acaaccttag agagttcact
attggtgatc ttaaatctgc cacaaggaac 240ttcagcaggt caggtatgat cggggaaggc
ggttttggtt gtgtcttctg gggaacaatc 300aagaacttag aagacccatc gaagaaaatc
gaagtcgcgg ttaaacagct cggcaaaaga 360gggttgcagg gtcataaaga atgggtgact
gaagtgaact ttctcggtgt agtcgagcat 420tcaaacttgg tgaagttgct gggacattgt
gcagaagacg atgaacgtgg aatccaaagg 480cttttggttt atgaatatat gccaaaccaa
agtgtcgagt tccatttatc tccgcggtca 540ccgacagtac ttacttggga cctcagattg
agaatagcac aagacgcagc tcgaggttta 600acataccttc atgaagaaat ggactttcag
ataatattcc gtgatttcaa gtcatccaac 660attctactag acgagaattg gacagcgaag
ctttcggatt tcggtttggc tcgcttaggt 720ccttcaccag gatccagcca tgtttctact
gatgtagtag gaacaatggg atacgcagct 780ccagagtata tccaaacggg tcgcctcacg
tcgaaaagcg atgtgtgggg atacggagtt 840ttcatctatg agctcattac aggaagaagg
ccactagacc ggaacaagcc taaaggagag 900cagaagcttt tagaatgggt gagaccttac
ttatccgaca caaggaggtt ccggctaata 960gtagacccga ggctcgaggg aaagtacatg
atcaagtcag tgcagaaact cgcggttgta 1020gccaaccttt gccttactag aaacgcaaag
gcgcgtccaa agatgagcga ggtgttagag 1080atggtgacaa agattgtgga agcttcatcg
cctgggaatg gtggcaagaa gccgcagctg 1140gttccactaa agagtcaaga aacttctaga
gtcgaggaag ggaagaataa gaaggttctt 1200gatggtgctg aaggaggttg gttagaaaag
ttgtggaacc caaagaatgt gagagcttgt 1260tga
126342420PRTarabidopsis thaliana 42Met
Lys Cys Phe Leu Phe Pro Leu Gly Asp Lys Lys Asp Glu Gln Arg 1
5 10 15 Ser Pro Lys Pro Val Ser
Pro Thr Ser Asn Phe Ser Asp Val Asn Lys 20
25 30 Ser Gly Ser Asp Phe Ser Pro Arg Asp Val
Ser Gly Thr Ser Thr Val 35 40
45 Ser Ser Thr Gly Arg Asn Ser Asn Thr Ser Met Ser Ala Arg
Glu Asn 50 55 60
Asn Leu Arg Glu Phe Thr Ile Gly Asp Leu Lys Ser Ala Thr Arg Asn 65
70 75 80 Phe Ser Arg Ser Gly
Met Ile Gly Glu Gly Gly Phe Gly Cys Val Phe 85
90 95 Trp Gly Thr Ile Lys Asn Leu Glu Asp Pro
Ser Lys Lys Ile Glu Val 100 105
110 Ala Val Lys Gln Leu Gly Lys Arg Gly Leu Gln Gly His Lys Glu
Trp 115 120 125 Val
Thr Glu Val Asn Phe Leu Gly Val Val Glu His Ser Asn Leu Val 130
135 140 Lys Leu Leu Gly His Cys
Ala Glu Asp Asp Glu Arg Gly Ile Gln Arg 145 150
155 160 Leu Leu Val Tyr Glu Tyr Met Pro Asn Gln Ser
Val Glu Phe His Leu 165 170
175 Ser Pro Arg Ser Pro Thr Val Leu Thr Trp Asp Leu Arg Leu Arg Ile
180 185 190 Ala Gln
Asp Ala Ala Arg Gly Leu Thr Tyr Leu His Glu Glu Met Asp 195
200 205 Phe Gln Ile Ile Phe Arg Asp
Phe Lys Ser Ser Asn Ile Leu Leu Asp 210 215
220 Glu Asn Trp Thr Ala Lys Leu Ser Asp Phe Gly Leu
Ala Arg Leu Gly 225 230 235
240 Pro Ser Pro Gly Ser Ser His Val Ser Thr Asp Val Val Gly Thr Met
245 250 255 Gly Tyr Ala
Ala Pro Glu Tyr Ile Gln Thr Gly Arg Leu Thr Ser Lys 260
265 270 Ser Asp Val Trp Gly Tyr Gly Val
Phe Ile Tyr Glu Leu Ile Thr Gly 275 280
285 Arg Arg Pro Leu Asp Arg Asn Lys Pro Lys Gly Glu Gln
Lys Leu Leu 290 295 300
Glu Trp Val Arg Pro Tyr Leu Ser Asp Thr Arg Arg Phe Arg Leu Ile 305
310 315 320 Val Asp Pro Arg
Leu Glu Gly Lys Tyr Met Ile Lys Ser Val Gln Lys 325
330 335 Leu Ala Val Val Ala Asn Leu Cys Leu
Thr Arg Asn Ala Lys Ala Arg 340 345
350 Pro Lys Met Ser Glu Val Leu Glu Met Val Thr Lys Ile Val
Glu Ala 355 360 365
Ser Ser Pro Gly Asn Gly Gly Lys Lys Pro Gln Leu Val Pro Leu Lys 370
375 380 Ser Gln Glu Thr Ser
Arg Val Glu Glu Gly Lys Asn Lys Lys Val Leu 385 390
395 400 Asp Gly Ala Glu Gly Gly Trp Leu Glu Lys
Leu Trp Asn Pro Lys Asn 405 410
415 Val Arg Ala Cys 420 431296DNAzea mays
43atgaggtgcc tgcctttctt gcatggagac accaaagaga aggatccagt cactaagtcg
60gcctctctac ggtccatgag cacaacatca acggagcgcg atgtccgctc cggttcagac
120ttcacctcct tgaatgtttc cgacatgagc gccgagtcga taaggaggac gcagtacccc
180agcttcactg accgcccgtc taacctgagg gtgttctcct ttgctgaact gaagagtgcc
240acccgcaact tcagccggtc tctcatggtt ggcgagggtg gctttggctg tgtgtacagg
300ggtgtcatca agacctccga tgaaccgaac gaacgaattg agatcgctgt taagcagttg
360aatcgtaaag gacttcaggg gcagaaggag tggttaacag agatgaatgt gcttggaatt
420gtggatcatc caaacctagt taaacttata ggctactgtg ctgaagatga tgagagggga
480gtacaacggc ttttagtgta cgaatatatg cctaatggaa gtgtggatga tcacttgtcg
540agtaggtcaa cttctactct gtcatggcca atgagactaa aagtagctct tgattctgct
600cggggactga agtatctgca tgaagaaatg gaattccagg ttattttccg ggacctgaaa
660acatctaaca ttttgttgga tgagaactgg aatgctaaac tgtcagactt tggtttggct
720aggcatggac cagcagaagg tctgacccat gtctccacag cggtggtcgg gactctaggc
780tacgcagctc cagagtacat gcagactggg cgcctgaccg cgaagagcga catatggagc
840tacggcgtcc tcctgtacga gctgatcaca ggccgccgcc ccatcgaccg gaaccgccca
900aagagcgagc agaagctcct ggactgggtg aagccgtaca tctcggacgt gaaacggttc
960cccatcatcg tcgacccgcg gctggagggg cactacaacc tcaagtccat gacgaagctg
1020tccagtgtgg cgaaccggtg cctggtccgg atgcccaagt ctcgccccaa gatgagcgag
1080gtgtacgaca tggtgcagaa gatcgtggac tgcgtgggga ccggcccgcc gcagcccccg
1140ctgctgcact accacggctc ggcctctgag cctggccctg gcgacaagcg cgccaggaaa
1200gggtcggtga agaggaggag gctctgggag ctcaggttcg gctgccggca catcgtgtgg
1260cgcggctgga agcctgcgat cgtgaaggac atctga
129644431PRTzea mays 44Met Arg Cys Leu Pro Phe Leu His Gly Asp Thr Lys
Glu Lys Asp Pro 1 5 10
15 Val Thr Lys Ser Ala Ser Leu Arg Ser Met Ser Thr Thr Ser Thr Glu
20 25 30 Arg Asp Val
Arg Ser Gly Ser Asp Phe Thr Ser Leu Asn Val Ser Asp 35
40 45 Met Ser Ala Glu Ser Ile Arg Arg
Thr Gln Tyr Pro Ser Phe Thr Asp 50 55
60 Arg Pro Ser Asn Leu Arg Val Phe Ser Phe Ala Glu Leu
Lys Ser Ala 65 70 75
80 Thr Arg Asn Phe Ser Arg Ser Leu Met Val Gly Glu Gly Gly Phe Gly
85 90 95 Cys Val Tyr Arg
Gly Val Ile Lys Thr Ser Asp Glu Pro Asn Glu Arg 100
105 110 Ile Glu Ile Ala Val Lys Gln Leu Asn
Arg Lys Gly Leu Gln Gly Gln 115 120
125 Lys Glu Trp Leu Thr Glu Met Asn Val Leu Gly Ile Val Asp
His Pro 130 135 140
Asn Leu Val Lys Leu Ile Gly Tyr Cys Ala Glu Asp Asp Glu Arg Gly 145
150 155 160 Val Gln Arg Leu Leu
Val Tyr Glu Tyr Met Pro Asn Gly Ser Val Asp 165
170 175 Asp His Leu Ser Ser Arg Ser Thr Ser Thr
Leu Ser Trp Pro Met Arg 180 185
190 Leu Lys Val Ala Leu Asp Ser Ala Arg Gly Leu Lys Tyr Leu His
Glu 195 200 205 Glu
Met Glu Phe Gln Val Ile Phe Arg Asp Leu Lys Thr Ser Asn Ile 210
215 220 Leu Leu Asp Glu Asn Trp
Asn Ala Lys Leu Ser Asp Phe Gly Leu Ala 225 230
235 240 Arg His Gly Pro Ala Glu Gly Leu Thr His Val
Ser Thr Ala Val Val 245 250
255 Gly Thr Leu Gly Tyr Ala Ala Pro Glu Tyr Met Gln Thr Gly Arg Leu
260 265 270 Thr Ala
Lys Ser Asp Ile Trp Ser Tyr Gly Val Leu Leu Tyr Glu Leu 275
280 285 Ile Thr Gly Arg Arg Pro Ile
Asp Arg Asn Arg Pro Lys Ser Glu Gln 290 295
300 Lys Leu Leu Asp Trp Val Lys Pro Tyr Ile Ser Asp
Val Lys Arg Phe 305 310 315
320 Pro Ile Ile Val Asp Pro Arg Leu Glu Gly His Tyr Asn Leu Lys Ser
325 330 335 Met Thr Lys
Leu Ser Ser Val Ala Asn Arg Cys Leu Val Arg Met Pro 340
345 350 Lys Ser Arg Pro Lys Met Ser Glu
Val Tyr Asp Met Val Gln Lys Ile 355 360
365 Val Asp Cys Val Gly Thr Gly Pro Pro Gln Pro Pro Leu
Leu His Tyr 370 375 380
His Gly Ser Ala Ser Glu Pro Gly Pro Gly Asp Lys Arg Ala Arg Lys 385
390 395 400 Gly Ser Val Lys
Arg Arg Arg Leu Trp Glu Leu Arg Phe Gly Cys Arg 405
410 415 His Ile Val Trp Arg Gly Trp Lys Pro
Ala Ile Val Lys Asp Ile 420 425
430 451287DNAglycine max 45atgaagtgtt ttccattctc gtatggagag
aaaaaagatg aaccgaaagg cttgcagttg 60cagtcaacat cgggtcgatc tgacaattcc
atgtgtgttg aggctgaggt tagaagatcc 120ggttctgagt taaattctca ggatgtttcg
gacaatggca gctcagaatc ccagaggagg 180aatgcaattc ccagtttgtc ccagagaccc
agcaacctca gagtgtttac tgtatctgaa 240ctgaaatcag ccaccaagaa tttcagtcgc
tctgttatga tcggagaggg tgggtttggg 300tgtgtctacc tgggattgat aagaagcgca
gaggactcct ccagaagaat tgaagttgca 360gttaaacaac ttagtaaaag aggaatgcag
ggccataggg aatgggtgac agaagtgaat 420gttctgggca ttgttgagca tcccaatctt
gtgaaactag tgggttactg tgctgatgat 480gatgaaagag gaatccagag gcttctaatt
tatgaataca tgccaaacag aagtgtggaa 540caccatttat ctcaccgatc agagactcct
ctcccatgga ctaggagatt aaaaatagct 600cgagatgcag ctcgtgggtt aacatacctg
catgaggaaa tggatttcca gataattttc 660agagatttca aatcttcaaa tatcctattg
gatgaacagt ggaatgcaaa gctatcagac 720tttgggttag caaggttggg accatcagat
ggactgactc atgtctcaac ggcggttgta 780ggaacaatgg gatatgccgc tcctgaatat
gttcaaaccg gacgtctaac ttcaaagaat 840gatgtatgga gctacggtgt cttcctttat
gaactcatca ctggtaggcg ccctttagat 900cgaaatcgcc ccaggcgtga gcagaagttg
ttggaatgga taaggccata cctatcagat 960gggaagaaat ttcaactaat attagatcca
agacttgata agaaacaagt cttcaagtca 1020gcccagagac tcgctatgat tgctaaccaa
tgcttggcaa aaaatcccaa gaatcgccca 1080aagatgagtg aggtattgga aatggtaaat
ggaatggtag aatcatcatc cagttctagt 1140ccacagttgc ccctgaggag tgtggtgaca
ttggaagctt cccaggatac tgaaacaaat 1200aacaagaaac gaaccatgga tcagaagctc
ggagaaagta attggtttgt taggatgtgg 1260agaccaaagc ttgtaagaac atgctga
128746428PRTglycine max 46Met Lys Cys
Phe Pro Phe Ser Tyr Gly Glu Lys Lys Asp Glu Pro Lys 1 5
10 15 Gly Leu Gln Leu Gln Ser Thr Ser
Gly Arg Ser Asp Asn Ser Met Cys 20 25
30 Val Glu Ala Glu Val Arg Arg Ser Gly Ser Glu Leu Asn
Ser Gln Asp 35 40 45
Val Ser Asp Asn Gly Ser Ser Glu Ser Gln Arg Arg Asn Ala Ile Pro 50
55 60 Ser Leu Ser Gln
Arg Pro Ser Asn Leu Arg Val Phe Thr Val Ser Glu 65 70
75 80 Leu Lys Ser Ala Thr Lys Asn Phe Ser
Arg Ser Val Met Ile Gly Glu 85 90
95 Gly Gly Phe Gly Cys Val Tyr Leu Gly Leu Ile Arg Ser Ala
Glu Asp 100 105 110
Ser Ser Arg Arg Ile Glu Val Ala Val Lys Gln Leu Ser Lys Arg Gly
115 120 125 Met Gln Gly His
Arg Glu Trp Val Thr Glu Val Asn Val Leu Gly Ile 130
135 140 Val Glu His Pro Asn Leu Val Lys
Leu Val Gly Tyr Cys Ala Asp Asp 145 150
155 160 Asp Glu Arg Gly Ile Gln Arg Leu Leu Ile Tyr Glu
Tyr Met Pro Asn 165 170
175 Arg Ser Val Glu His His Leu Ser His Arg Ser Glu Thr Pro Leu Pro
180 185 190 Trp Thr Arg
Arg Leu Lys Ile Ala Arg Asp Ala Ala Arg Gly Leu Thr 195
200 205 Tyr Leu His Glu Glu Met Asp Phe
Gln Ile Ile Phe Arg Asp Phe Lys 210 215
220 Ser Ser Asn Ile Leu Leu Asp Glu Gln Trp Asn Ala Lys
Leu Ser Asp 225 230 235
240 Phe Gly Leu Ala Arg Leu Gly Pro Ser Asp Gly Leu Thr His Val Ser
245 250 255 Thr Ala Val Val
Gly Thr Met Gly Tyr Ala Ala Pro Glu Tyr Val Gln 260
265 270 Thr Gly Arg Leu Thr Ser Lys Asn Asp
Val Trp Ser Tyr Gly Val Phe 275 280
285 Leu Tyr Glu Leu Ile Thr Gly Arg Arg Pro Leu Asp Arg Asn
Arg Pro 290 295 300
Arg Arg Glu Gln Lys Leu Leu Glu Trp Ile Arg Pro Tyr Leu Ser Asp 305
310 315 320 Gly Lys Lys Phe Gln
Leu Ile Leu Asp Pro Arg Leu Asp Lys Lys Gln 325
330 335 Val Phe Lys Ser Ala Gln Arg Leu Ala Met
Ile Ala Asn Gln Cys Leu 340 345
350 Ala Lys Asn Pro Lys Asn Arg Pro Lys Met Ser Glu Val Leu Glu
Met 355 360 365 Val
Asn Gly Met Val Glu Ser Ser Ser Ser Ser Ser Pro Gln Leu Pro 370
375 380 Leu Arg Ser Val Val Thr
Leu Glu Ala Ser Gln Asp Thr Glu Thr Asn 385 390
395 400 Asn Lys Lys Arg Thr Met Asp Gln Lys Leu Gly
Glu Ser Asn Trp Phe 405 410
415 Val Arg Met Trp Arg Pro Lys Leu Val Arg Thr Cys 420
425 471284DNAoryza sativa 47atgaggtgcc
tgcctttctt gcatggagat tccaaagagg aggatcccgt caacaagtcg 60gcttctgtcc
ggtcgttgag cacaacatcg acggagcggg atgtccggtc cggctccgac 120ttcaactcct
tgaatgtctc tgacatgagt gccgaatcaa tacggaggac acagtatccc 180agcttcactg
atcggcccag taacctcagg gtgttctctt tctctgagct gaagaatgcc 240actcgcaatt
ttagccggtc tcttatggtt ggtgagggtg ggtttggatg tgtgtatagg 300ggtgtcatca
agaattccga tgaaccaact gagcgcaccg agattgctgt taaacagctg 360aatcgcaaag
gacttcaggg gcagaaagaa tggttaacag aactgaatgt gcttgggatt 420gtagagcatc
caaacctcgt caaactaatt ggctactgcg ctgaagatga tgaaaggggc 480gtacagcgtc
tcctagtata cgaatacatg cctaatggaa gcgtggatga tcacttgtca 540agtaggtcaa
attcaactct atcatggcca atgagactaa aagtagctct ggacgctgct 600cggggactga
agtatctgca tgaagagatg gaatttcagg ttatcttccg tgacctaaaa 660acatctaaca
ttctgttaga tgagaactgg aatgcaaagt tgtctgactt tggattggct 720aggcatggac
catcagaagg cctgacccat gtctctacag cggtcgtggg aactcttggg 780tatgcagctc
cggagtacat gcagaccgga cgcctcactg ccaagagtga catatggggc 840tatggtgtgc
tcctttatga gctcatcacc ggccgccgtc ccattgaccg gaaccgccca 900aagggtgagc
agaagctcct ggattgggtg aaaccataca tatctgatat caagcggttc 960cccatcatca
tagacccacg gctagagggg cactacaacc tcaagtccat gacaaagctg 1020gctagtgtgg
cgaaccgctg tctcgtccgg ctaccaaagt cgcgcccaaa gatgagtgag 1080gtgtatgaga
tggttcagaa gattgtggcc agcattgaga ccggcacacc acagcctcct 1140ctgcactacc
atgggtcggt ttctgaaccg ggctcaaagc ggccaaagaa ggggtcactg 1200aagagaaggt
tccaagaatt caaattcggt tgccggcaga ttgtatggcg gagctggaag 1260cctgagatca
taaagacttg ctga
128448428PRToryza sativa 48Met Met Arg Cys Leu Pro Phe Leu His Gly Asp
Ser Lys Glu Glu Asp 1 5 10
15 Pro Val Asn Lys Ser Ala Ser Val Arg Ser Leu Ser Thr Thr Ser Thr
20 25 30 Glu Arg
Asp Val Arg Ser Gly Ser Asp Phe Asn Ser Leu Asn Val Ser 35
40 45 Asp Met Ser Ala Glu Ser Ile
Arg Arg Thr Gln Tyr Pro Ser Phe Thr 50 55
60 Asp Arg Pro Ser Asn Leu Arg Val Phe Ser Phe Ser
Glu Leu Lys Asn 65 70 75
80 Ala Thr Arg Asn Phe Ser Arg Ser Leu Met Val Gly Glu Gly Gly Phe
85 90 95 Gly Cys Val
Tyr Arg Gly Val Ile Lys Asn Ser Asp Glu Pro Thr Glu 100
105 110 Arg Thr Glu Ile Ala Val Lys Gln
Leu Asn Arg Lys Gly Leu Gln Gly 115 120
125 Gln Lys Glu Trp Leu Thr Glu Leu Asn Val Leu Gly Ile
Val Glu His 130 135 140
Pro Asn Leu Val Lys Leu Ile Gly Tyr Cys Ala Glu Asp Asp Glu Arg 145
150 155 160 Gly Val Gln Arg
Leu Leu Val Tyr Glu Tyr Met Pro Asn Gly Ser Val 165
170 175 Asp Asp His Leu Ser Ser Arg Ser Asn
Ser Thr Leu Ser Trp Pro Met 180 185
190 Arg Leu Lys Val Ala Leu Asp Ala Ala Arg Gly Leu Lys Tyr
Leu His 195 200 205
Glu Glu Met Glu Phe Gln Val Ile Phe Arg Asp Leu Lys Thr Ser Asn 210
215 220 Ile Leu Leu Asp Glu
Asn Trp Asn Ala Lys Leu Ser Asp Phe Gly Leu 225 230
235 240 Ala Arg His Gly Pro Ser Glu Gly Leu Thr
His Val Ser Thr Ala Val 245 250
255 Val Gly Thr Leu Gly Tyr Ala Ala Pro Glu Tyr Met Gln Thr Gly
Arg 260 265 270 Leu
Thr Ala Lys Ser Asp Ile Trp Gly Tyr Gly Val Leu Leu Tyr Glu 275
280 285 Leu Ile Thr Gly Arg Arg
Pro Ile Asp Arg Asn Arg Pro Lys Gly Glu 290 295
300 Gln Lys Leu Leu Asp Trp Val Lys Pro Tyr Ile
Ser Asp Ile Lys Arg 305 310 315
320 Phe Pro Ile Ile Ile Asp Pro Arg Leu Glu Gly His Tyr Asn Leu Lys
325 330 335 Ser Met
Thr Lys Leu Ala Ser Val Ala Asn Arg Cys Leu Val Arg Leu 340
345 350 Pro Lys Ser Arg Pro Lys Met
Ser Glu Val Tyr Glu Met Val Gln Lys 355 360
365 Ile Val Ala Ser Ile Glu Thr Gly Thr Pro Gln Pro
Pro Leu His Tyr 370 375 380
His Gly Ser Val Ser Glu Pro Gly Ser Lys Arg Pro Lys Lys Gly Ser 385
390 395 400 Leu Lys Arg
Arg Phe Gln Glu Phe Lys Phe Gly Cys Arg Gln Ile Val 405
410 415 Trp Arg Ser Trp Lys Pro Glu Ile
Ile Lys Thr Cys 420 425
491041DNAGossypium hirsutum 49acagtttctg agcttaaatc tgcaaccaag aactttagcc
gctctttcat gctcggagag 60ggtggatttg gctgtgttta caagggttct ctcaagagtc
ctgaagatcc gtctgaaaag 120attgaagtag cagtgaaaca gcttggtaaa agggggttgc
agggccacaa ggagtgggtg 180actgaagtaa atgtccttgg tgtggttgag catccgaatc
ttgtgaagct agttggttac 240tgtgctgaag acgatgaaag aggaatccaa cggcttttga
tatatgaata tatgcctaat 300agaagtgtgg aaaaccattt atctgtgcgg tcagaaacaa
ctctttcctg ggcaatgaga 360ttgaaaatag cccaagatgc tgctcgtggg ttagcatacc
tacatgaagg aatggagttc 420cagatcatct tcagggattt taaatcatca aatatccttc
tagatgagca atggaatgca 480aagctctctg actttggatt agccaggttg ggcccttcag
aaggattaac tcatatctca 540acagcggttg ttgggacaat gggatatgcg gctcctgaat
acatccagac aggacgttta 600acatccaaga ttgatgtgtg gagctatggg gtcttcctct
atgaactcat tactggcagg 660cgcccctttg acaaaaaccg tcccaagaat gagcaaaggc
tattggaatg ggtaaagcca 720tacctatctg ataggaaatt ccagttgata ttggacccta
gactgaaagg gaaataccaa 780ctcaagtctg ctcaaaggct tgcggttgtg gccaaccgat
gcttagtcag aaacccaaag 840tcacgcccta agatgagtga ggttttagaa atggtgaatc
ggattgtgga agcatcatca 900gcaggaccca gaactcctga accaccattg aatgatgtct
ctctggaaac tgctagggaa 960cgtaaaagaa ggattataga ttttagaagc ggtgaagagt
ttgtttggtc atggactcca 1020aagctcataa gaccatgcta a
104150346PRTGossypium hirsutum 50Thr Val Ser Glu
Leu Lys Ser Ala Thr Lys Asn Phe Ser Arg Ser Phe 1 5
10 15 Met Leu Gly Glu Gly Gly Phe Gly Cys
Val Tyr Lys Gly Ser Leu Lys 20 25
30 Ser Pro Glu Asp Pro Ser Glu Lys Ile Glu Val Ala Val Lys
Gln Leu 35 40 45
Gly Lys Arg Gly Leu Gln Gly His Lys Glu Trp Val Thr Glu Val Asn 50
55 60 Val Leu Gly Val Val
Glu His Pro Asn Leu Val Lys Leu Val Gly Tyr 65 70
75 80 Cys Ala Glu Asp Asp Glu Arg Gly Ile Gln
Arg Leu Leu Ile Tyr Glu 85 90
95 Tyr Met Pro Asn Arg Ser Val Glu Asn His Leu Ser Val Arg Ser
Glu 100 105 110 Thr
Thr Leu Ser Trp Ala Met Arg Leu Lys Ile Ala Gln Asp Ala Ala 115
120 125 Arg Gly Leu Ala Tyr Leu
His Glu Gly Met Glu Phe Gln Ile Ile Phe 130 135
140 Arg Asp Phe Lys Ser Ser Asn Ile Leu Leu Asp
Glu Gln Trp Asn Ala 145 150 155
160 Lys Leu Ser Asp Phe Gly Leu Ala Arg Leu Gly Pro Ser Glu Gly Leu
165 170 175 Thr His
Ile Ser Thr Ala Val Val Gly Thr Met Gly Tyr Ala Ala Pro 180
185 190 Glu Tyr Ile Gln Thr Gly Arg
Leu Thr Ser Lys Ile Asp Val Trp Ser 195 200
205 Tyr Gly Val Phe Leu Tyr Glu Leu Ile Thr Gly Arg
Arg Pro Phe Asp 210 215 220
Lys Asn Arg Pro Lys Asn Glu Gln Arg Leu Leu Glu Trp Val Lys Pro 225
230 235 240 Tyr Leu Ser
Asp Arg Lys Phe Gln Leu Ile Leu Asp Pro Arg Leu Lys 245
250 255 Gly Lys Tyr Gln Leu Lys Ser Ala
Gln Arg Leu Ala Val Val Ala Asn 260 265
270 Arg Cys Leu Val Arg Asn Pro Lys Ser Arg Pro Lys Met
Ser Glu Val 275 280 285
Leu Glu Met Val Asn Arg Ile Val Glu Ala Ser Ser Ala Gly Pro Arg 290
295 300 Thr Pro Glu Pro
Pro Leu Asn Asp Val Ser Leu Glu Thr Ala Arg Glu 305 310
315 320 Arg Lys Arg Arg Ile Ile Asp Phe Arg
Ser Gly Glu Glu Phe Val Trp 325 330
335 Ser Trp Thr Pro Lys Leu Ile Arg Pro Cys 340
345 5130DNAArtificial SequenceNG6 Forward Primer
51tcgagctagc atgaagcttg tggagaagac
305229DNAArtificial SequenceNG6 Reverse Primer 52cgacgagctc ttacctgatg
gaacaagag 295322DNAArtificial
SequenceNG21 Forward Primer 53atggtgagtg acaagcatgt ag
225420DNAArtificial SequenceNG21 Reverse Primer
54tcacttgccc gtgatgaatg
205521DNAArtificial SequenceNG24 Forward Primer 55atgggttatc tctcttgcaa c
215620DNAArtificial
SequenceNG24 Reverse Primer 56tcagtatctc ttccgcgacg
205720DNAArtificial SequenceNG28 Forward Primer
57atgggcatgg aagctttgag
205820DNAArtificial SequenceNG28 Reverse Primer 58tcaaaatgga gtttcggcgt
205920DNAArtificial
SequenceNG32 Forward Primer 59atgaaatgct tcttattccc
206020DNAArtificial SequenceNG32 Reverse Primer
60tcaacaagct ctcacattct
206118DNAArtificial SequenceNG6 Forward Primer 61tgtgcccgga gccctacc
186222DNAArtificial
SequenceNG6 Reverse Primer 62ctttcagtgc catgcggatt tt
226321DNAArtificial SequenceNG21 Forward Primer
63ggcacaagtc ccgtcatcac c
216422DNAArtificial SequenceNG21 Reverse Primer 64tccccaatcc cttcttttcc
ta 226520DNAArtificial
SequenceNG24 Forward Primer 65gccgccgtca agagaacaac
206621DNAArtificial SequenceNG24 Reverse Primer
66ctccggtggt caacgcagta a
216722DNAArtificial SequenceNG28 Forward Primer 67tgttgttgtg gcctcgttgt
ta 226822DNAArtificial
SequenceNG28 Reverse Primer 68ctttccttca ccgccttctt tc
226921DNAArtificial SequenceNG32 Forward Primer
69aagctttcgg atttcggttt g
217022DNAArtificial SequenceNG32 Reverse Primer 70tggccttctt cctgtaatga
gc 227117DNAArtificial
SequenceACTIN Forward Primer 71cccgctatgt atgtcgc
177218DNAArtificial SequenceACTIN Reverse
Primer 72aaggtcaaga cggaggat
18731971DNAArabidopsis thaliana 73atgatcgtta atttctcttt cttcctcctt
ctcttcgtct ccgtcttcgt ttcctctgct 60gattctaaag cgaccatttc aatttcccct
aatgctctca atcgatctgg cgattccgtt 120gtgatacaat ggtccggtgt cgattctccg
tcagatctcg attggttagg actctactcg 180ccgccggagt ctcctaatga tcactttatt
ggttacaaat tcctcaatga atcgtccact 240tggaaagatg gtttcggttc gatttctctt
cctttaacca atctccgatc aaattacaca 300ttccggatct tccgttggag cgaatccgag
attgatccga aacataagga tcatgatcag 360aatcctttac caggaactaa acatcttcta
gctgaatcgg agcagctgac tttcggatcc 420ggtgttggta tgccggagca gatccatttg
tcgttcacaa atatggttaa cacgatgcgt 480gttatgtttg tagctggaga tggtgaagaa
cgttttgtta gatacggtga atcgaaggat 540ttgttaggta attccgcggc ggcgcgtggg
atgaggtacg agagagagca catgtgtgat 600tcgccggcga attccactat tggttggaga
gatcctggtt ggatttttga taccgtcatg 660aagaatttga atgatggcgt tagatactat
tatcaggttg ggagtgattc taagggatgg 720agtgagatcc atagctacat tgctcgagat
gtgactgcag aagaaaccgt agctttcatg 780tttggagata tgggttgtgc tacaccatac
acgacattta tccgcacaca agatgagagc 840atatctacag tgaagtggat cctccgtgac
attgaagctc ttggtgataa gccagctatg 900atttcacaca ttggagatat aagttatgct
cgtggttact cgtgggtatg ggatgagttc 960tttgctcagg ttgagcctat tgcctcgaca
gttccttacc atgtttgcat tggtaaccat 1020gagtatgatt tctctactca gccgtggaaa
cctgattggg cagcttctat ttatggaaac 1080gatggtggtg gcgaatgtgg tgtgccgtat
agcttgaagt ttaacatgcc tgggaattct 1140tcagagtcta caggaatgaa agctcctccg
acaaggaatt tatattattc ttatgatatg 1200ggaacggtcc atttcgttta tatctccaca
gagacgaatt ttcttaaagg aggtagtcaa 1260tatgaattca taaagcgaga tctagagtct
gtagacagga agaaaacacc gtttgttgtt 1320gtgcaaggac atagaccaat gtacactacg
agcaacgagg ttagagacac tatgattcga 1380caaaagatgg ttgagcatct agaacctttg
tttgtgaaaa acaatgtcac acttgctcta 1440tggggacatg ttcatagata cgaaaggttt
tgtcccataa gcaacaacac ttgcggcaca 1500cagtggcaag gaaatccggt tcatcttgtg
atcggtatgg ctggtcaaga ttggcaaccg 1560atttggcagc ctagaccaaa ccatccagat
cttcctatat tccctcagcc tgaacaatca 1620atgtatcgta caggtgagtt tggttacact
cgtttagttg caaacaaaga aaagctcact 1680gtttcttttg tgggtaatca cgatggcgaa
gttcatgata ctgttgagat gttagcatct 1740ggggtagtaa tcagtgggag caaagagagt
actaaaatcc caaatctgaa aaccgttcct 1800gcttctgcta cacttatggg aaaatcagaa
tctaatgctt tgtggtatgc caaaggagca 1860ggcttgatgg ttgtgggtgt gcttttaggg
ttcattatcg ggttttttac ccggggaaag 1920aaatcttcgt ctggaaaccg ttggatccca
gtcaagaacg aggagacata a 197174656PRTArabidopsis thaliana 74Met
Ile Val Asn Phe Ser Phe Phe Leu Leu Leu Phe Val Ser Val Phe 1
5 10 15 Val Ser Ser Ala Asp Ser
Lys Ala Thr Ile Ser Ile Ser Pro Asn Ala 20
25 30 Leu Asn Arg Ser Gly Asp Ser Val Val Ile
Gln Trp Ser Gly Val Asp 35 40
45 Ser Pro Ser Asp Leu Asp Trp Leu Gly Leu Tyr Ser Pro Pro
Glu Ser 50 55 60
Pro Asn Asp His Phe Ile Gly Tyr Lys Phe Leu Asn Glu Ser Ser Thr 65
70 75 80 Trp Lys Asp Gly Phe
Gly Ser Ile Ser Leu Pro Leu Thr Asn Leu Arg 85
90 95 Ser Asn Tyr Thr Phe Arg Ile Phe Arg Trp
Ser Glu Ser Glu Ile Asp 100 105
110 Pro Lys His Lys Asp His Asp Gln Asn Pro Leu Pro Gly Thr Lys
His 115 120 125 Leu
Leu Ala Glu Ser Glu Gln Leu Thr Phe Gly Ser Gly Val Gly Met 130
135 140 Pro Glu Gln Ile His Leu
Ser Phe Thr Asn Met Val Asn Thr Met Arg 145 150
155 160 Val Met Phe Val Ala Gly Asp Gly Glu Glu Arg
Phe Val Arg Tyr Gly 165 170
175 Glu Ser Lys Asp Leu Leu Gly Asn Ser Ala Ala Ala Arg Gly Met Arg
180 185 190 Tyr Glu
Arg Glu His Met Cys Asp Ser Pro Ala Asn Ser Thr Ile Gly 195
200 205 Trp Arg Asp Pro Gly Trp Ile
Phe Asp Thr Val Met Lys Asn Leu Asn 210 215
220 Asp Gly Val Arg Tyr Tyr Tyr Gln Val Gly Ser Asp
Ser Lys Gly Trp 225 230 235
240 Ser Glu Ile His Ser Tyr Ile Ala Arg Asp Val Thr Ala Glu Glu Thr
245 250 255 Val Ala Phe
Met Phe Gly Asp Met Gly Cys Ala Thr Pro Tyr Thr Thr 260
265 270 Phe Ile Arg Thr Gln Asp Glu Ser
Ile Ser Thr Val Lys Trp Ile Leu 275 280
285 Arg Asp Ile Glu Ala Leu Gly Asp Lys Pro Ala Met Ile
Ser His Ile 290 295 300
Gly Asp Ile Ser Tyr Ala Arg Gly Tyr Ser Trp Val Trp Asp Glu Phe 305
310 315 320 Phe Ala Gln Val
Glu Pro Ile Ala Ser Thr Val Pro Tyr His Val Cys 325
330 335 Ile Gly Asn His Glu Tyr Asp Phe Ser
Thr Gln Pro Trp Lys Pro Asp 340 345
350 Trp Ala Ala Ser Ile Tyr Gly Asn Asp Gly Gly Gly Glu Cys
Gly Val 355 360 365
Pro Tyr Ser Leu Lys Phe Asn Met Pro Gly Asn Ser Ser Glu Ser Thr 370
375 380 Gly Met Lys Ala Pro
Pro Thr Arg Asn Leu Tyr Tyr Ser Tyr Asp Met 385 390
395 400 Gly Thr Val His Phe Val Tyr Ile Ser Thr
Glu Thr Asn Phe Leu Lys 405 410
415 Gly Gly Ser Gln Tyr Glu Phe Ile Lys Arg Asp Leu Glu Ser Val
Asp 420 425 430 Arg
Lys Lys Thr Pro Phe Val Val Val Gln Gly His Arg Pro Met Tyr 435
440 445 Thr Thr Ser Asn Glu Val
Arg Asp Thr Met Ile Arg Gln Lys Met Val 450 455
460 Glu His Leu Glu Pro Leu Phe Val Lys Asn Asn
Val Thr Leu Ala Leu 465 470 475
480 Trp Gly His Val His Arg Tyr Glu Arg Phe Cys Pro Ile Ser Asn Asn
485 490 495 Thr Cys
Gly Thr Gln Trp Gln Gly Asn Pro Val His Leu Val Ile Gly 500
505 510 Met Ala Gly Gln Asp Trp Gln
Pro Ile Trp Gln Pro Arg Pro Asn His 515 520
525 Pro Asp Leu Pro Ile Phe Pro Gln Pro Glu Gln Ser
Met Tyr Arg Thr 530 535 540
Gly Glu Phe Gly Tyr Thr Arg Leu Val Ala Asn Lys Glu Lys Leu Thr 545
550 555 560 Val Ser Phe
Val Gly Asn His Asp Gly Glu Val His Asp Thr Val Glu 565
570 575 Met Leu Ala Ser Gly Val Val Ile
Ser Gly Ser Lys Glu Ser Thr Lys 580 585
590 Ile Pro Asn Leu Lys Thr Val Pro Ala Ser Ala Thr Leu
Met Gly Lys 595 600 605
Ser Glu Ser Asn Ala Leu Trp Tyr Ala Lys Gly Ala Gly Leu Met Val 610
615 620 Val Gly Val Leu
Leu Gly Phe Ile Ile Gly Phe Phe Thr Arg Gly Lys 625 630
635 640 Lys Ser Ser Ser Gly Asn Arg Trp Ile
Pro Val Lys Asn Glu Glu Thr 645 650
655 758PRTArtificial sequencemotifs 75Gly Ile Phe Arg Ser
Gly Phe Pro 1 5 768PRTArtificial
sequencemotifs 76Tyr Leu Cys Pro Glu Pro Tyr Pro 1 5
778PRTArtificial sequencemotifs 77Lys Glu Pro Phe Val Xaa Ile Pro
1 5 7810PRTArtificial sequencemotifs 78His
Cys Xaa Arg Gly Lys His Arg Thr Gly 1 5
10 7910PRTArtificial sequenceNG21 motif 79Asp Asn Trp Asp Asp Ala Xaa
Gly Tyr Tyr 1 5 10 8012PRTArtificial
sequenceNG21 motif 80Tyr Arg Asn His Leu Cys Leu Val Phe Glu Ser Leu 1
5 10 8115PRTArtificial sequenceNG21
motif 81Val Leu His Cys Asp Ile Lys Pro Asp Asn Met Leu Val Asn Glu 1
5 10 15 8214PRTArtificial
sequenceNG21 motif 82Thr Pro Tyr Leu Val Ser Arg Phe Tyr Arg Xaa Pro Glu
Ile 1 5 10
838PRTArtificial sequenceNG24 motif 83Val Arg His Arg Asp Xaa Lys Ser 1
5 849PRTArtificial sequenceNG24 motif 84Gly Thr
Leu Xaa Gly Tyr Leu Asp Pro 1 5
8512PRTArtificial SequenceNG24 motif 85Asp Val Xaa Ser Xaa Gly Xaa Leu
Leu Leu Glu Ile 1 5 10
868PRTArtificial sequenceNG28 motif 86Arg Arg His Lys Ile Ala Leu Gly 1
5 876PRTArtificial sequenceNG28 motif 87Tyr Xaa
Ala Pro Glu Leu 1 5 8811PRTArtificial sequenceNG28
motif 88Asp Val Tyr Ala Phe Gly Ile Leu Leu Leu Glu 1 5
10 898PRTArtificial sequenceNG32 motif 89Cys Ala Xaa
Asp Asp Glu Arg Gly 1 5 9010PRTArtificial
sequenceNG32 motif 90Ala Lys Leu Ser Asp Phe Gly Leu Ala Arg 1
5 10 918PRTArtificial SequenceNG32 motif 91Tyr Glu
Leu Ile Thr Gly Arg Xaa 1 5 927PRTArtificial
sequenceNG32 motif 92Arg Pro Lys Met Ser Glu Val 1 5
93648DNABrassica napus 93atgaagctag ttgagaatac gccggcggcg actgacaagt
tcaccaccac ggaggaggag 60gacggtgaag acgcctgccg cacgatcgag gtcgtcgaga
gaaacgtgtt tcaggctcag 120ttcgatgaag ctgctgatgc ggttgaggag cttaacctta
taccgccgct caacttctct 180atggtggata acggaatatt ccgttctgga ttccctgatc
cggctaactt ctccttcctc 240cagactctcg gactccgctc aattatttat ctgtgtccgg
agccttaccc agagagtaac 300atccagttcc tcaaatccaa tgggattact cttttccagt
ttggcattga aggcaataag 360gaaccatttg tgattattcc agaccagaaa atccgcaagg
cactcaatgt ccttttagat 420gagaaaaacc atccggttct gattcattgt aagcgaggca
agcatcgtac tggttgtctt 480gtgggttgct tgagaaaact tcagaaatgg tgtttgactt
cgatatttga cgagtaccag 540cgatttgcag cagctaaagc tagagtttca gatcaaagat
tcatggagat attcgacgtc 600tccagcttca gtcatgttcc gatgtctttc tcttgttcca
gcaggtaa 64894215PRTBrassica napus 94Met Lys Leu Val Glu
Asn Thr Pro Ala Ala Thr Asp Lys Phe Thr Thr 1 5
10 15 Thr Glu Glu Glu Asp Gly Glu Asp Ala Cys
Arg Thr Ile Glu Val Val 20 25
30 Glu Arg Asn Val Phe Gln Ala Gln Phe Asp Glu Ala Ala Asp Ala
Val 35 40 45 Glu
Glu Leu Asn Leu Ile Pro Pro Leu Asn Phe Ser Met Val Asp Asn 50
55 60 Gly Ile Phe Arg Ser Gly
Phe Pro Asp Pro Ala Asn Phe Ser Phe Leu 65 70
75 80 Gln Thr Leu Gly Leu Arg Ser Ile Ile Tyr Leu
Cys Pro Glu Pro Tyr 85 90
95 Pro Glu Ser Asn Ile Gln Phe Leu Lys Ser Asn Gly Ile Thr Leu Phe
100 105 110 Gln Phe
Gly Ile Glu Gly Asn Lys Glu Pro Phe Val Ile Ile Pro Asp 115
120 125 Gln Lys Ile Arg Lys Ala Leu
Asn Val Leu Leu Asp Glu Lys Asn His 130 135
140 Pro Val Leu Ile His Cys Lys Arg Gly Lys His Arg
Thr Gly Cys Leu 145 150 155
160 Val Gly Cys Leu Arg Lys Leu Gln Lys Trp Cys Leu Thr Ser Ile Phe
165 170 175 Asp Glu Tyr
Gln Arg Phe Ala Ala Ala Lys Ala Arg Val Ser Asp Gln 180
185 190 Arg Phe Met Glu Ile Phe Asp Val
Ser Ser Phe Ser His Val Pro Met 195 200
205 Ser Phe Ser Cys Ser Ser Arg 210
215 952325DNABrassica napus 95gtggtgtccg gtgcattccc ggcggccctt gaaaatccgg
aggacaagaa agaatctggg 60ggtccttcgg agagagttgg taaaagaagt tatgataatg
gcaggagttc gttttcttcg 120gagaactctg aagagaagta taagagtagt aaccggtctc
ttacggagag ccggcagtat 180aacgaagtcc gtgcgcggag tagatctaag tcaagggttg
ttgcggagga tgagttttcg 240gtcagaggaa gacatcgcga ctctagtagg gagtatcgtc
atgacagagt tgactcaagg 300aggagcgagg gacgtgggcg atatgaagga tatgacaggg
aatatactcg agaagacgtg 360gaaagagaaa gaagtaagga gagggatatg gacagggaag
gaagcattcg agatagggat 420tcagaaggaa gtaaacggag agagagggat attgatcgga
ggagggaaag agaacgagaa 480gaaaggaggg agatagaggc tgaccgtgaa aggcggaaag
ataaggaacg ggagcgcagc 540attgataggg ataggagaag ggagagggaa ggacgagata
gagacaatga aagaggtggg 600agcgtcgata gggaaaggag aagggagagg gaaggagatt
atttacgaga cagagacaat 660agaaggggta ggagtagaga cagaaccaga tatgatagcc
gagagaggat gagagaaaag 720gaaagggaga gtgacaaaga tagagaaatc caggctgata
aggagaagca taaaagtgtt 780gaagtggaca acggtgaaag gtcgaaatat gagaatgatc
aagatgataa tgacaaagaa 840tttatatgga aatctccgga agaaatagaa gaagaagaat
taaataaaat cagggagagc 900atagagaaat ttaaaaagaa gcccgagcag caaagtgaac
ttatttcgca ggataaggag 960atagatttcg ttcaagaaag cagtgctcca gattcggctt
cttttgcagt tgttacagat 1020gctaatgctg gtgcagccaa agctaagtcg gacttcgacc
ctgtagttag tgatgttgct 1080aaaacctcat taacagctgg tgggccacct aatatgtttg
gaatttcaaa ctcggagaaa 1140actcaagctc cagcagggct tggcgaaggt agcccaaaga
gtgaaagatc agctgacatg 1200tttcatgatg atatatttgg agagtcccca gctgctaatc
aaaaagtgga tcacatgcga 1260gggaaaggtg atggtgttcc aatggtaagg agcgggcttc
atgacaattg ggatgatgcg 1320gagggttatt acagctatca gttcggcgag ctaattgacg
gcagatatga agtcattgct 1380actcatggaa aaggcgtttt ctctactgtc gttcgtgcga
aagatttaag agctggacca 1440gctgaacctg acgaagttgc tataaaaatt attcgtaaca
acgagacgat gcataaagct 1500ggccagactg aggttcagat attgaagaag ctggctggtg
ctgaccgaga tgacaagcgg 1560cactgtgttc gtttgctttc aagtttcaag taccggaatc
acctttgctt ggtgtttgag 1620tcgctccatt taaatcttcg agagctcttg aagaagtttg
gccgtaacat tggcctcaaa 1680ctgtctgctg ttaggtcgta ttcaaagcag cttttcattg
cccttaaaca tctgaagaat 1740tgtggggttc ttcactgcga tataaaacct gacaacatgc
tggtgaatga gaacaaaacc 1800gtgttgaagc tttgcgactt tggtaatgca atgtttgctg
gaaaaaacga agtcacgcca 1860tatctcgtta gtcgctttta cagatcccct gaaatcattc
tggggctggc ttatgaccat 1920ccgcttgata tatggtcggt tggctgctgt ctatatgagc
tttattgcgg gaaagttctt 1980ttccctggcg ccacaaataa tgatatgtta cgccttcata
tggaactgaa aggccttttc 2040cccaaaaaga tgcttcgtaa gggagcattt attgatcagc
actttgatca cgacttgaac 2100ttttacgcta cagaggagga cactgttagt gggaagatga
tgaagagaat gattttaaat 2160gtaaagccaa aagattttgg ttcaattata aagggttacc
ctggtgagga tcccaagatg 2220ttagctcatt tcagggatct cttagacaag atgttcatcc
ttgatccaga gaagagactg 2280actgtgtcac aggcattagc tcacccattt atcactggca
agtga 232596774PRTBrassica napus 96Met Val Ser Gly Ala
Phe Pro Ala Ala Leu Glu Asn Pro Glu Asp Lys 1 5
10 15 Lys Glu Ser Gly Gly Pro Ser Glu Arg Val
Gly Lys Arg Ser Tyr Asp 20 25
30 Asn Gly Arg Ser Ser Phe Ser Ser Glu Asn Ser Glu Glu Lys Tyr
Lys 35 40 45 Ser
Ser Asn Arg Ser Leu Thr Glu Ser Arg Gln Tyr Asn Glu Val Arg 50
55 60 Ala Arg Ser Arg Ser Lys
Ser Arg Val Val Ala Glu Asp Glu Phe Ser 65 70
75 80 Val Arg Gly Arg His Arg Asp Ser Ser Arg Glu
Tyr Arg His Asp Arg 85 90
95 Val Asp Ser Arg Arg Ser Glu Gly Arg Gly Arg Tyr Glu Gly Tyr Asp
100 105 110 Arg Glu
Tyr Thr Arg Glu Asp Val Glu Arg Glu Arg Ser Lys Glu Arg 115
120 125 Asp Met Asp Arg Glu Gly Ser
Ile Arg Asp Arg Asp Ser Glu Gly Ser 130 135
140 Lys Arg Arg Glu Arg Asp Ile Asp Arg Arg Arg Glu
Arg Glu Arg Glu 145 150 155
160 Glu Arg Arg Glu Ile Glu Ala Asp Arg Glu Arg Arg Lys Asp Lys Glu
165 170 175 Arg Glu Arg
Ser Ile Asp Arg Asp Arg Arg Arg Glu Arg Glu Gly Arg 180
185 190 Asp Arg Asp Asn Glu Arg Gly Gly
Ser Val Asp Arg Glu Arg Arg Arg 195 200
205 Glu Arg Glu Gly Asp Tyr Leu Arg Asp Arg Asp Asn Arg
Arg Gly Arg 210 215 220
Ser Arg Asp Arg Thr Arg Tyr Asp Ser Arg Glu Arg Met Arg Glu Lys 225
230 235 240 Glu Arg Glu Ser
Asp Lys Asp Arg Glu Ile Gln Ala Asp Lys Glu Lys 245
250 255 His Lys Ser Val Glu Val Asp Asn Gly
Glu Arg Ser Lys Tyr Glu Asn 260 265
270 Asp Gln Asp Asp Asn Asp Lys Glu Phe Ile Trp Lys Ser Pro
Glu Glu 275 280 285
Ile Glu Glu Glu Glu Leu Asn Lys Ile Arg Glu Ser Ile Glu Lys Phe 290
295 300 Lys Lys Lys Pro Glu
Gln Gln Ser Glu Leu Ile Ser Gln Asp Lys Glu 305 310
315 320 Ile Asp Phe Val Gln Glu Ser Ser Ala Pro
Asp Ser Ala Ser Phe Ala 325 330
335 Val Val Thr Asp Ala Asn Ala Gly Ala Ala Lys Ala Lys Ser Asp
Phe 340 345 350 Asp
Pro Val Val Ser Asp Val Ala Lys Thr Ser Leu Thr Ala Gly Gly 355
360 365 Pro Pro Asn Met Phe Gly
Ile Ser Asn Ser Glu Lys Thr Gln Ala Pro 370 375
380 Ala Gly Leu Gly Glu Gly Ser Pro Lys Ser Glu
Arg Ser Ala Asp Met 385 390 395
400 Phe His Asp Asp Ile Phe Gly Glu Ser Pro Ala Ala Asn Gln Lys Val
405 410 415 Asp His
Met Arg Gly Lys Gly Asp Gly Val Pro Met Val Arg Ser Gly 420
425 430 Leu His Asp Asn Trp Asp Asp
Ala Glu Gly Tyr Tyr Ser Tyr Gln Phe 435 440
445 Gly Glu Leu Ile Asp Gly Arg Tyr Glu Val Ile Ala
Thr His Gly Lys 450 455 460
Gly Val Phe Ser Thr Val Val Arg Ala Lys Asp Leu Arg Ala Gly Pro 465
470 475 480 Ala Glu Pro
Asp Glu Val Ala Ile Lys Ile Ile Arg Asn Asn Glu Thr 485
490 495 Met His Lys Ala Gly Gln Thr Glu
Val Gln Ile Leu Lys Lys Leu Ala 500 505
510 Gly Ala Asp Arg Asp Asp Lys Arg His Cys Val Arg Leu
Leu Ser Ser 515 520 525
Phe Lys Tyr Arg Asn His Leu Cys Leu Val Phe Glu Ser Leu His Leu 530
535 540 Asn Leu Arg Glu
Leu Leu Lys Lys Phe Gly Arg Asn Ile Gly Leu Lys 545 550
555 560 Leu Ser Ala Val Arg Ser Tyr Ser Lys
Gln Leu Phe Ile Ala Leu Lys 565 570
575 His Leu Lys Asn Cys Gly Val Leu His Cys Asp Ile Lys Pro
Asp Asn 580 585 590
Met Leu Val Asn Glu Asn Lys Thr Val Leu Lys Leu Cys Asp Phe Gly
595 600 605 Asn Ala Met Phe
Ala Gly Lys Asn Glu Val Thr Pro Tyr Leu Val Ser 610
615 620 Arg Phe Tyr Arg Ser Pro Glu Ile
Ile Leu Gly Leu Ala Tyr Asp His 625 630
635 640 Pro Leu Asp Ile Trp Ser Val Gly Cys Cys Leu Tyr
Glu Leu Tyr Cys 645 650
655 Gly Lys Val Leu Phe Pro Gly Ala Thr Asn Asn Asp Met Leu Arg Leu
660 665 670 His Met Glu
Leu Lys Gly Leu Phe Pro Lys Lys Met Leu Arg Lys Gly 675
680 685 Ala Phe Ile Asp Gln His Phe Asp
His Asp Leu Asn Phe Tyr Ala Thr 690 695
700 Glu Glu Asp Thr Val Ser Gly Lys Met Met Lys Arg Met
Ile Leu Asn 705 710 715
720 Val Lys Pro Lys Asp Phe Gly Ser Ile Ile Lys Gly Tyr Pro Gly Glu
725 730 735 Asp Pro Lys Met
Leu Ala His Phe Arg Asp Leu Leu Asp Lys Met Phe 740
745 750 Ile Leu Asp Pro Glu Lys Arg Leu Thr
Val Ser Gln Ala Leu Ala His 755 760
765 Pro Phe Ile Thr Gly Lys 770
971387DNABrassica napus 97atgcagctac ctattgcaac tgcagagatt ggctttgcgt
tgcttctaac tgcagctgta 60tcaatatcag cggttttata cgttcggtat aggctgaggc
attgtaggtg ctcagagagt 120gatgcaaggt cttctaaaga ctcagcgttt accaaagata
acgaccgtcc ggatcttgat 180aagttgcaga agcgcagaag ggctagagtg ttcacctacg
aggagctgga gaaagctgca 240gaagggttca aagaagagtc aatagtaggg aaagggagct
tctcatgtgt gtacaaaggt 300gtgctgagag atggaaccac tgtcgctgtg aagaaggcca
taatgtcatc agacaaacag 360aagaactcaa acgagttccg caccgagctt gatctgttgt
caagactcaa ccatgctcat 420ctccttagcc ttcttggcta ctgcgaagaa ggaggagaga
ggcttcttgt ttacgagttt 480atggcgcatg gctcactcta caaccatctt cacggtaaga
acaaggcctt gaaagagcag 540ctagattggg ttaaacgagt caccattgct gtccaagcag
ctagaggaat cgagtacttg 600catggctacg cttgtcctcc tgtcatccac cgtgatatca
aatcatcaaa catacttata 660gacgaagaac acaacgctag agtagctgac tttggtctct
ccttgcttgg tcctgttgat 720agcggctctc ctttagcaga gctgccagct ggcactctcg
gttaccttga tcctgagtac 780tatagactac actatctcac aaccaagtct gatgtctaca
gcttcggagt cttgcttctc 840gagatcctga gcggaagaaa agctattgac atgcactatg
aagaagggaa catagtggaa 900tgggcggttc ctttgatcaa agctggagat attacatcaa
tcttggaccc ggtcttgaaa 960caaccaaccg agatagaagc tctgaggagg atagtgagcg
tggcttgcaa atgcgtgagg 1020atgagaggca aagacagacc gtcaatggat aaagtgacaa
catcactgga gagagctctc 1080gcgcagctga tggggaaccc gagcagcgag cagccgatat
taccgacaga agtggttctt 1140gggagcagca ggatgcacaa gaagtcgtgg aggatcggtt
cggagaacac tgagtttaga 1200ggcgggtcgt ggataacttt ccctagcgtg acgtcatcac
agaggaggaa gtcttcagct 1260tctgaaggtg atgtggcgga ggaggtggag gatgaaggaa
ggaagcaaca agaggcgttg 1320aggagtcttg aagaggagat aggaccagct tctcctggac
agagcttgtt cttgcatcat 1380aatttct
138798462PRTBrassica napus 98Met Gln Leu Pro Ile
Ala Thr Ala Glu Ile Gly Phe Ala Leu Leu Leu 1 5
10 15 Thr Ala Ala Val Ser Ile Ser Ala Val Leu
Tyr Val Arg Tyr Arg Leu 20 25
30 Arg His Cys Arg Cys Ser Glu Ser Asp Ala Arg Ser Ser Lys Asp
Ser 35 40 45 Ala
Phe Thr Lys Asp Asn Asp Arg Pro Asp Leu Asp Lys Leu Gln Lys 50
55 60 Arg Arg Arg Ala Arg Val
Phe Thr Tyr Glu Glu Leu Glu Lys Ala Ala 65 70
75 80 Glu Gly Phe Lys Glu Glu Ser Ile Val Gly Lys
Gly Ser Phe Ser Cys 85 90
95 Val Tyr Lys Gly Val Leu Arg Asp Gly Thr Thr Val Ala Val Lys Lys
100 105 110 Ala Ile
Met Ser Ser Asp Lys Gln Lys Asn Ser Asn Glu Phe Arg Thr 115
120 125 Glu Leu Asp Leu Leu Ser Arg
Leu Asn His Ala His Leu Leu Ser Leu 130 135
140 Leu Gly Tyr Cys Glu Glu Gly Gly Glu Arg Leu Leu
Val Tyr Glu Phe 145 150 155
160 Met Ala His Gly Ser Leu Tyr Asn His Leu His Gly Lys Asn Lys Ala
165 170 175 Leu Lys Glu
Gln Leu Asp Trp Val Lys Arg Val Thr Ile Ala Val Gln 180
185 190 Ala Ala Arg Gly Ile Glu Tyr Leu
His Gly Tyr Ala Cys Pro Pro Val 195 200
205 Ile His Arg Asp Ile Lys Ser Ser Asn Ile Leu Ile Asp
Glu Glu His 210 215 220
Asn Ala Arg Val Ala Asp Phe Gly Leu Ser Leu Leu Gly Pro Val Asp 225
230 235 240 Ser Gly Ser Pro
Leu Ala Glu Leu Pro Ala Gly Thr Leu Gly Tyr Leu 245
250 255 Asp Pro Glu Tyr Tyr Arg Leu His Tyr
Leu Thr Thr Lys Ser Asp Val 260 265
270 Tyr Ser Phe Gly Val Leu Leu Leu Glu Ile Leu Ser Gly Arg
Lys Ala 275 280 285
Ile Asp Met His Tyr Glu Glu Gly Asn Ile Val Glu Trp Ala Val Pro 290
295 300 Leu Ile Lys Ala Gly
Asp Ile Thr Ser Ile Leu Asp Pro Val Leu Lys 305 310
315 320 Gln Pro Thr Glu Ile Glu Ala Leu Arg Arg
Ile Val Ser Val Ala Cys 325 330
335 Lys Cys Val Arg Met Arg Gly Lys Asp Arg Pro Ser Met Asp Lys
Val 340 345 350 Thr
Thr Ser Leu Glu Arg Ala Leu Ala Gln Leu Met Gly Asn Pro Ser 355
360 365 Ser Glu Gln Pro Ile Leu
Pro Thr Glu Val Val Leu Gly Ser Ser Arg 370 375
380 Met His Lys Lys Ser Trp Arg Ile Gly Ser Glu
Asn Thr Glu Phe Arg 385 390 395
400 Gly Gly Ser Trp Ile Thr Phe Pro Ser Val Thr Ser Ser Gln Arg Arg
405 410 415 Lys Ser
Ser Ala Ser Glu Gly Asp Val Ala Glu Glu Val Glu Asp Glu 420
425 430 Gly Arg Lys Gln Gln Glu Ala
Leu Arg Ser Leu Glu Glu Glu Ile Gly 435 440
445 Pro Ala Ser Pro Gly Gln Ser Leu Phe Leu His His
Asn Phe 450 455 460
99612DNABrassica napus 99gccaggagac acaagattgc gttgggaata gcgagaggac
ttgcttatct tcacaccgga 60caagaagctc ccatcatcca cggcaatata agatcgaaaa
acgtgctggt ggacgacttc 120ttcttcgcta ggctgactga gtttgggctt gataagatca
tggtgcaagc ggtggcggat 180gagatagtct cgcaggcgaa atcagacgga tacaaggcgc
ctgagcttca caagatgaag 240aaatgcaacc cgagaagcga tgtttacgcc tttgggatcc
ttctcctgga gatactgatg 300ggcaagaagc ctgggaagag tgggaggaac ggtggtgagt
atgttgatct accttctttg 360gttaaagccg cggtgttgga ggagacgacg atggaggttt
tcgacttgga ggcgatgaaa 420gggatcagga gtccgatgga ggaaggtttg gttcatgcgt
tgaagctggc gatgggatgc 480tgtgctcctg ttacgacggt tagaccgagt atggaagagg
ttgtgaagca gttggaagag 540aacaggccga ggaatagatc agcgttgtat agccctacgg
aaacgaggag cgacgctgag 600acaccatgct ga
612100203PRTBrassica napus 100Ala Arg Arg His Lys
Ile Ala Leu Gly Ile Ala Arg Gly Leu Ala Tyr 1 5
10 15 Leu His Thr Gly Gln Glu Ala Pro Ile Ile
His Gly Asn Ile Arg Ser 20 25
30 Lys Asn Val Leu Val Asp Asp Phe Phe Phe Ala Arg Leu Thr Glu
Phe 35 40 45 Gly
Leu Asp Lys Ile Met Val Gln Ala Val Ala Asp Glu Ile Val Ser 50
55 60 Gln Ala Lys Ser Asp Gly
Tyr Lys Ala Pro Glu Leu His Lys Met Lys 65 70
75 80 Lys Cys Asn Pro Arg Ser Asp Val Tyr Ala Phe
Gly Ile Leu Leu Leu 85 90
95 Glu Ile Leu Met Gly Lys Lys Pro Gly Lys Ser Gly Arg Asn Gly Gly
100 105 110 Glu Tyr
Val Asp Leu Pro Ser Leu Val Lys Ala Ala Val Leu Glu Glu 115
120 125 Thr Thr Met Glu Val Phe Asp
Leu Glu Ala Met Lys Gly Ile Arg Ser 130 135
140 Pro Met Glu Glu Gly Leu Val His Ala Leu Lys Leu
Ala Met Gly Cys 145 150 155
160 Cys Ala Pro Val Thr Thr Val Arg Pro Ser Met Glu Glu Val Val Lys
165 170 175 Gln Leu Glu
Glu Asn Arg Pro Arg Asn Arg Ser Ala Leu Tyr Ser Pro 180
185 190 Thr Glu Thr Arg Ser Asp Ala Glu
Thr Pro Cys 195 200
1011278DNABrassica napus 101atgaagtgtt tcaagttctc tagtggtgac aagaaagaag
aacacaacaa gactcccaag 60tctgtctcac tgacctctaa cttctccgac cgcgacataa
accgaagcgg gtcggatttc 120aactctcgag acgcctctgg gacgagcacg gagtcgtcca
tggggaggaa gaactcgtac 180ccttcaatgt ctgctagaga aagtaatctc agagagttca
gcgttactga tctcaaggct 240gcgactaaga actttagccg ctctgttatg attggagaag
gagggttcgg ttgtgtcttc 300aggggaacag tgagggactt ggaagatccg tcgattaaaa
tcgaagtcgc ggttaagcag 360ctcggtaaaa gagggttgca ggggcataag gaatgggtca
cggaagtgaa ctttcttggt 420gtggttgagc attcaaactt ggtgaagttg cttggttact
gtgcagaaga tgatgaacgt 480gggatccaac ggcttttggt ttatgaatac atgccaaacc
gaagcgttga gtcccactta 540tcccctcgct cactcacagt ccttacttgg gatctaaggc
tgagaatcgc tcaagatgca 600gctcgtggtt taacatacct gcatgaacaa atggagtttc
agataatatt cagggacttt 660aagtcctcga acattctctt ggatgaggac tggaaagcaa
agctctctga ctttggcctg 720gctcgtttag gtccatctga aggactaact catgttacta
ctgatgttgt aggtacaatg 780gcttatgcag ctcctgagta tattcaaact ggtcgtctca
catcaaaaag cgatgtgtgg 840ggttatggag tgtttatcta cgagctcatc acagggagga
aaccagttga taggaacaaa 900cctaagggag agcagaagct tctagaatgg gtgagacctt
atctatcaga cacaaggaag 960ttcaagctca tattagaccc gaggctagaa gggaagtacc
ctctcaaatc agttcagaag 1020ctagcggttg tggccaacag gtgtttagtt agaaacccaa
aggcacgtcc caagatgagt 1080gaagtgctgg agatggtgaa caagattgtg gaagcgcctt
catgtagcgg tactagcccg 1140cagctagttc cgctgcaggg tctggagact tccagagacg
ctggaggagg gaaaaagaag 1200aggggtttag agaatggtgg tggtgaagga ggttggtttg
gtaagttatg gaacccaaag 1260acaataagag cttgttga
1278102425PRTBrassica napus 102Met Lys Cys Phe Lys
Phe Ser Ser Gly Asp Lys Lys Glu Glu His Asn 1 5
10 15 Lys Thr Pro Lys Ser Val Ser Leu Thr Ser
Asn Phe Ser Asp Arg Asp 20 25
30 Ile Asn Arg Ser Gly Ser Asp Phe Asn Ser Arg Asp Ala Ser Gly
Thr 35 40 45 Ser
Thr Glu Ser Ser Met Gly Arg Lys Asn Ser Tyr Pro Ser Met Ser 50
55 60 Ala Arg Glu Ser Asn Leu
Arg Glu Phe Ser Val Thr Asp Leu Lys Ala 65 70
75 80 Ala Thr Lys Asn Phe Ser Arg Ser Val Met Ile
Gly Glu Gly Gly Phe 85 90
95 Gly Cys Val Phe Arg Gly Thr Val Arg Asp Leu Glu Asp Pro Ser Ile
100 105 110 Lys Ile
Glu Val Ala Val Lys Gln Leu Gly Lys Arg Gly Leu Gln Gly 115
120 125 His Lys Glu Trp Val Thr Glu
Val Asn Phe Leu Gly Val Val Glu His 130 135
140 Ser Asn Leu Val Lys Leu Leu Gly Tyr Cys Ala Glu
Asp Asp Glu Arg 145 150 155
160 Gly Ile Gln Arg Leu Leu Val Tyr Glu Tyr Met Pro Asn Arg Ser Val
165 170 175 Glu Ser His
Leu Ser Pro Arg Ser Leu Thr Val Leu Thr Trp Asp Leu 180
185 190 Arg Leu Arg Ile Ala Gln Asp Ala
Ala Arg Gly Leu Thr Tyr Leu His 195 200
205 Glu Gln Met Glu Phe Gln Ile Ile Phe Arg Asp Phe Lys
Ser Ser Asn 210 215 220
Ile Leu Leu Asp Glu Asp Trp Lys Ala Lys Leu Ser Asp Phe Gly Leu 225
230 235 240 Ala Arg Leu Gly
Pro Ser Glu Gly Leu Thr His Val Thr Thr Asp Val 245
250 255 Val Gly Thr Met Ala Tyr Ala Ala Pro
Glu Tyr Ile Gln Thr Gly Arg 260 265
270 Leu Thr Ser Lys Ser Asp Val Trp Gly Tyr Gly Val Phe Ile
Tyr Glu 275 280 285
Leu Ile Thr Gly Arg Lys Pro Val Asp Arg Asn Lys Pro Lys Gly Glu 290
295 300 Gln Lys Leu Leu Glu
Trp Val Arg Pro Tyr Leu Ser Asp Thr Arg Lys 305 310
315 320 Phe Lys Leu Ile Leu Asp Pro Arg Leu Glu
Gly Lys Tyr Pro Leu Lys 325 330
335 Ser Val Gln Lys Leu Ala Val Val Ala Asn Arg Cys Leu Val Arg
Asn 340 345 350 Pro
Lys Ala Arg Pro Lys Met Ser Glu Val Leu Glu Met Val Asn Lys 355
360 365 Ile Val Glu Ala Pro Ser
Cys Ser Gly Thr Ser Pro Gln Leu Val Pro 370 375
380 Leu Gln Gly Leu Glu Thr Ser Arg Asp Ala Gly
Gly Gly Lys Lys Lys 385 390 395
400 Arg Gly Leu Glu Asn Gly Gly Gly Glu Gly Gly Trp Phe Gly Lys Leu
405 410 415 Trp Asn
Pro Lys Thr Ile Arg Ala Cys 420 425
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