Patent application title: AXMI-192 FAMILY OF PESTICIDAL GENES AND METHODS FOR THEIR USE
Inventors:
Kimberly S. Sampson (Durham, NC, US)
Daniel J. Tomso (Bahama, NC, US)
Rong Guo (Cary, NC, US)
Rong Guo (Cary, NC, US)
IPC8 Class: AC07K200FI
USPC Class:
514 45
Class name: Designated organic active ingredient containing (doai) peptide (e.g., protein, etc.) containing doai insect destroying or inhibiting
Publication date: 2013-11-14
Patent application number: 20130303440
Abstract:
Compositions and methods for conferring pesticidal activity to bacteria,
plants, plant cells, tissues and seeds are provided. Compositions
comprising a coding sequence for a toxin polypeptide are provided. The
coding sequences can be used in DNA constructs or expression cassettes
for transformation and expression in plants and bacteria. Compositions
also comprise transformed bacteria, plants, plant cells, tissues, and
seeds. In particular, isolated toxin nucleic acid molecules are provided.
Additionally, amino acid sequences corresponding to the polynucleotides
are encompassed, and antibodies specifically binding to those amino acid
sequences. In particular, the present invention provides for isolated
nucleic acid molecules comprising nucleotide sequences encoding the amino
acid sequence shown in SEQ ID NO:28-62, or the nucleotide sequence set
forth in SEQ ID NO:1-27, as well as variants and fragments thereof.Claims:
1. A recombinant polypeptide with pesticidal activity, selected from the
group consisting of: a) a polypeptide comprising the amino acid sequence
of SEQ ID NO:34 or 35; b) a polypeptide comprising an amino acid sequence
having at least 95% sequence identity to the amino acid sequence of SEQ
ID NO:34 or 35, wherein said amino acid sequence has pesticidal activity;
and c) a polypeptide that is encoded by the nucleotide sequence of SEQ ID
NO:7.
2. The polypeptide of claim 1 further comprising heterologous amino acid sequences.
3. A composition comprising the polypeptide of claim 1.
4. The composition of claim 3, wherein said composition is selected from the group consisting of a powder, dust, pellet, granule, spray, emulsion, colloid, and solution.
5. The composition of claim 3, wherein said composition is prepared by desiccation, lyophilization, homogenization, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of Bacillus thuringiensis cells.
6. The composition of claim 3, comprising from about 1% to about 99% by weight of said polypeptide.
7. A method for controlling a lepidopteran, coleopteran, or nematode pest population comprising contacting said population with an pesticidally-effective amount of the polypeptide of claim 1.
8. A method for killing a lepidopteran, coleopteran, or nematode pest, comprising contacting said pest with, or feeding to said pest, an pesticidally-effective amount of the polypeptide of claim 1.
Description:
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. patent application Ser. No. 12/846,900, filed Jul. 30, 2010, which claims the benefit of U.S. Provisional Application Ser. No. 61/230,659, filed Jul. 31, 2009, the contents of which are herein incorporated by reference in their entirety.
REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY
[0002] The official copy of the sequence listing is submitted electronically via EFS-Web as an ASCII formatted sequence listing with a file named "APA067US01DSEQLIST.txt", created on May 14, 2013, and having a size of 273 kilobytes and is filed concurrently with the specification. The sequence listing contained in this ASCII formatted document is part of the specification and is herein incorporated by reference in its entirety.
FIELD OF THE INVENTION
[0003] This invention relates to the field of molecular biology. Provided are novel genes that encode pesticidal proteins. These proteins and the nucleic acid sequences that encode them are useful in preparing pesticidal formulations and in the production of transgenic pest-resistant plants.
BACKGROUND OF THE INVENTION
[0004] Bacillus thuringiensis is a Gram-positive spore forming soil bacterium characterized by its ability to produce crystalline inclusions that are specifically toxic to certain orders and species of insects, but are harmless to plants and other non-targeted organisms. For this reason, compositions including Bacillus thuringiensis strains or their insecticidal proteins can be used as environmentally-acceptable insecticides to control agricultural insect pests or insect vectors for a variety of human or animal diseases.
[0005] Crystal (Cry) proteins (delta-endotoxins) from Bacillus thuringiensis have potent insecticidal activity against predominantly Lepidopteran, Dipteran, and Coleopteran larvae. These proteins also have shown activity against Hymenoptera, Homoptera, Phthiraptera, Mallophaga, and Acari pest orders, as well as other invertebrate orders such as Nemathelminthes, Platyhelminthes, and Sarcomastigorphora (Feitelson (1993) The Bacillus Thuringiensis family tree. In Advanced Engineered Pesticides, Marcel Dekker, Inc., New York, N.Y.) These proteins were originally classified as CryI to CryV based primarily on their insecticidal activity. The major classes were Lepidoptera-specific (I), Lepidoptera- and Diptera-specific (II), Coleoptera-specific (III), Diptera-specific (IV), and nematode-specific (V) and (VI). The proteins were further classified into subfamilies; more highly related proteins within each family were assigned divisional letters such as Cry1A, Cry1B, Cry1C, etc. Even more closely related proteins within each division were given names such as Cry1C1, Cry1C2, etc.
[0006] A new nomenclature was recently described for the Cry genes based upon amino acid sequence homology rather than insect target specificity (Crickmore et al. (1998) Microbiol. Mol. Biol. Rev. 62:807-813). In the new classification, each toxin is assigned a unique name incorporating a primary rank (an Arabic number), a secondary rank (an uppercase letter), a tertiary rank (a lowercase letter), and a quaternary rank (another Arabic number). In the new classification, Roman numerals have been exchanged for Arabic numerals in the primary rank. Proteins with less than 45% sequence identity have different primary ranks, and the criteria for secondary and tertiary ranks are 78% and 95%, respectively.
[0007] The crystal protein does not exhibit insecticidal activity until it has been ingested and solubilized in the insect midgut. The ingested protoxin is hydrolyzed by proteases in the insect digestive tract to an active toxic molecule. (Hofte and Whiteley (1989) Microbiol. Rev. 53:242-255). This toxin binds to apical brush border receptors in the midgut of the target larvae and inserts into the apical membrane creating ion channels or pores, resulting in larval death.
[0008] Delta-endotoxins generally have five conserved sequence domains, and three conserved structural domains (see, for example, de Maagd et al. (2001) Trends Genetics 17:193-199). The first conserved structural domain consists of seven alpha helices and is involved in membrane insertion and pore formation. Domain II consists of three beta-sheets arranged in a Greek key configuration, and domain III consists of two antiparallel beta-sheets in "jelly-roll" formation (de Maagd et al., 2001, supra). Domains II and III are involved in receptor recognition and binding, and are therefore considered determinants of toxin specificity.
[0009] Aside from delta-endotoxins, there are several other known classes of pesticidal protein toxins. The VIP1/VIP2 toxins (see, for example, U.S. Pat. No. 5,770,696) are binary pesticidal toxins that exhibit strong activity on insects by a mechanism believed to involve receptor-mediated endocytosis followed by cellular toxification, similar to the mode of action of other binary ("A/B") toxins. A/B toxins such as VIP, C2, CDT, CST, or the B. anthracis edema and lethal toxins initially interact with target cells via a specific, receptor-mediated binding of "B" components as monomers. These monomers then form homoheptamers. The "B" heptamer-receptor complex then acts as a docking platform that subsequently binds and allows the translocation of an enzymatic "A" component(s) into the cytosol via receptor-mediated endocytosis. Once inside the cell's cytosol, "A" components inhibit normal cell function by, for example, ADP-ribosylation of G-actin, or increasing intracellular levels of cyclic AMP (cAMP). See Barth et al. (2004) Microbiol Mol Biol Rev 68:373-402.
[0010] The intensive use of B. thuringiensis-based insecticides has already given rise to resistance in field populations of the diamondback moth, Plutella xylostella (Ferre and Van Rie (2002) Annu. Rev. Entomol. 47:501-533). The most common mechanism of resistance is the reduction of binding of the toxin to its specific midgut receptor(s). This may also confer cross-resistance to other toxins that share the same receptor (Ferre and Van Rie (2002)).
SUMMARY OF INVENTION
[0011] Compositions and methods for conferring pest resistance to bacteria, plants, plant cells, tissues and seeds are provided. Compositions include nucleic acid molecules encoding sequences for toxin polypeptides, vectors comprising those nucleic acid molecules, and host cells comprising the vectors. Compositions also include the polypeptide sequences of the toxin, and antibodies to those polypeptides. The nucleotide sequences can be used in DNA constructs or expression cassettes for transformation and expression in organisms, including microorganisms and plants. The nucleotide or amino acid sequences may be synthetic sequences that have been designed for expression in an organism including, but not limited to, a microorganism or a plant. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds.
[0012] In particular, isolated or recombinant nucleic acid molecules corresponding to toxin nucleic acid sequences are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed. In particular, the present invention provides for an isolated nucleic acid molecule comprising a nucleotide sequence encoding the amino acid sequence shown in any of SEQ ID NO:28-62, or a nucleotide sequence set forth in any of SEQ ID NO:1-27, as well as variants and fragments thereof. Nucleotide sequences that are complementary to a nucleotide sequence of the invention, or that hybridize to a sequence of the invention are also encompassed.
[0013] The compositions and methods of the invention are useful for the production of organisms with pesticide resistance, specifically bacteria and plants. These organisms and compositions derived from them are desirable for agricultural purposes. The compositions of the invention are also useful for generating altered or improved toxin proteins that have pesticidal activity, or for detecting the presence of toxin proteins or nucleic acids in products or organisms.
DETAILED DESCRIPTION
[0014] The present invention is drawn to compositions and methods for regulating pest resistance or tolerance in organisms, particularly plants or plant cells. By "resistance" is intended that the pest (e.g., insect) is killed upon ingestion or other contact with the polypeptides of the invention. By "tolerance" is intended an impairment or reduction in the movement, feeding, reproduction, or other functions of the pest. The methods involve transforming organisms with a nucleotide sequence encoding a pesticidal protein of the invention. In particular, the nucleotide sequences of the invention are useful for preparing plants and microorganisms that possess pesticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are pesticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest, as probes for the isolation of other homologous (or partially homologous) genes, and for the generation of altered pesticidal proteins by methods known in the art, such as domain swapping or DNA shuffling. The proteins find use in controlling or killing lepidopteran, coleopteran, dipteran, and nematode pest populations and for producing compositions with pesticidal activity.
[0015] By "pesticidal toxin" or "pesticidal protein" is intended a toxin that has toxic activity against one or more pests, including, but not limited to, members of the Lepidoptera, Diptera, and Coleoptera orders, or the Nematoda phylum, or a protein that has homology to such a protein. Pesticidal proteins have been isolated from organisms including, for example, Bacillus sp., Clostridium bifermentans and Paenibacillus popilliae. Pesticidal proteins include amino acid sequences deduced from the full-length nucleotide sequences disclosed herein, and amino acid sequences that are shorter than the full-length sequences, either due to the use of an alternate downstream start site, or due to processing that produces a shorter protein having pesticidal activity. Processing may occur in the organism the protein is expressed in, or in the pest after ingestion of the protein.
[0016] In various embodiments, the sequences disclosed herein have homology to delta-endotoxin proteins. Delta-endotoxins include proteins identified as cry1 through cry53, cytl and cyt2, and Cyt-like toxin. There are currently over 250 known species of delta-endotoxins with a wide range of specificities and toxicities. For an expansive list see Crickmore et al. (1998), Microbiol. Mol. Biol. Rev. 62:807-813, and for regular updates see Crickmore et al. (2003) "Bacillus thuringiensis toxin nomenclature," at www.biols.susx.ac.uk/Home/Neil_Crickmore/Bt/index. In some embodiments, the delta-endotoxin sequences disclosed herein include the nucleotide sequences set forth in any of SEQ ID NO:1-27, the amino acid sequences set forth in any of SEQ ID NO:28-62, as well as variants and fragments thereof.
[0017] Thus, provided herein are novel isolated or recombinant nucleotide sequences that confer pesticidal activity. Also provided are the amino acid sequences of the pesticidal proteins. The protein resulting from translation of this gene allows cells to control or kill pests that ingest it.
Isolated Nucleic Acid Molecules, and Variants and Fragments Thereof
[0018] One aspect of the invention pertains to isolated or recombinant nucleic acid molecules comprising nucleotide sequences encoding pesticidal proteins and polypeptides or biologically active portions thereof, as well as nucleic acid molecules sufficient for use as hybridization probes to identify nucleic acid molecules encoding proteins with regions of sequence homology. As used herein, the term "nucleic acid molecule" is intended to include DNA molecules (e.g., recombinant DNA, cDNA or genomic DNA) and RNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated using nucleotide analogs. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA.
[0019] An "isolated" nucleic acid sequence (or DNA) is used herein to refer to a nucleic acid sequence (or DNA) that is no longer in its natural environment, for example in an in vitro or in a recombinant bacterial or plant host cell. In some embodiments, an "isolated" nucleic acid is free of sequences (preferably protein encoding sequences) that naturally flank the nucleic acid (i.e., sequences located at the 5' and 3' ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For purposes of the invention, "isolated" when used to refer to nucleic acid molecules excludes isolated chromosomes. For example, in various embodiments, the isolated nucleic acid molecule encoding a pesticidal protein can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb of nucleotide sequences that naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived. A pesticidal protein that is substantially free of cellular material includes preparations of protein having less than about 30%, 20%, 10%, or 5% (by dry weight) of non-pesticidal protein (also referred to herein as a "contaminating protein").
[0020] Nucleotide sequences encoding the proteins of the present invention include the sequence set forth in SEQ ID NO:1-27, and variants, fragments, and complements thereof. By "complement" is intended a nucleotide sequence that is sufficiently complementary to a given nucleotide sequence such that it can hybridize to the given nucleotide sequence to thereby form a stable duplex. The corresponding amino acid sequence for the toxin protein encoded by this nucleotide sequence are set forth in SEQ ID NO:28-62.
[0021] Nucleic acid molecules that are fragments of these nucleotide sequences encoding pesticidal proteins are also encompassed by the present invention. By "fragment" is intended a portion of the nucleotide sequence encoding a pesticidal protein. A fragment of a nucleotide sequence may encode a biologically active portion of a pesticidal protein, or it may be a fragment that can be used as a hybridization probe or PCR primer using methods disclosed below. Nucleic acid molecules that are fragments of a nucleotide sequence encoding a pesticidal protein comprise at least about 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1350, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2500, 3000, 3500, 4000, 4500, 5000 contiguous nucleotides, or up to the number of nucleotides present in a full-length nucleotide sequence encoding a pesticidal protein disclosed herein, depending upon the intended use. By "contiguous" nucleotides is intended nucleotide residues that are immediately adjacent to one another. Fragments of the nucleotide sequences of the present invention will encode protein fragments that retain the biological activity of the pesticidal protein and, hence, retain pesticidal activity. By "retains activity" is intended that the fragment will have at least about 30%, at least about 50%, at least about 70%, 80%, 90%, 95% or higher of the pesticidal activity of the pesticidal protein. In one embodiment, the pesticidal activity is coleoptericidal activity. In another embodiment, the pesticidal activity is lepidoptericidal activity. In another embodiment, the pesticidal activity is nematocidal activity. In another embodiment, the pesticidal activity is diptericidal activity. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety.
[0022] A fragment of a nucleotide sequence that encodes a biologically active portion of a pesticidal protein of the invention will encode at least about 15, 25, 30, 50, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100 contiguous amino acids, or up to the total number of amino acids present in a full-length pesticidal protein of the invention. In some embodiments, the fragment is a C-terminal truncation of at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, 450, 500, 650, 600 or more amino acids relative to the amino acid sequences of the invention.
[0023] Preferred pesticidal proteins of the present invention are encoded by a nucleotide sequence sufficiently identical to the nucleotide sequence of SEQ ID NO:1-27. By "sufficiently identical" is intended an amino acid or nucleotide sequence that has at least about 60% or 65% sequence identity, about 70% or 75% sequence identity, about 80% or 85% sequence identity, about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity compared to a reference sequence using one of the alignment programs described herein using standard parameters. One of skill in the art will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by two nucleotide sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning, and the like.
[0024] To determine the percent identity of two amino acid sequences or of two nucleic acids, the sequences are aligned for optimal comparison purposes. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., percent identity=number of identical positions/total number of positions (e.g., overlapping positions)×100). In another embodiment, the comparison is across the entirety of the reference sequence (i.e., across the entirety of any of SEQ ID NO:1-61) The percent identity between two sequences can be determined using techniques similar to those described below, with or without allowing gaps. In calculating percent identity, typically exact matches are counted.
[0025] The determination of percent identity between two sequences can be accomplished using a mathematical algorithm. A nonlimiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA 87:2264, modified as in Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877. Such an algorithm is incorporated into the BLASTN and BLASTX programs of Altschul et al. (1990) J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed with the BLASTN program, score=100, wordlength=12, to obtain nucleotide sequences homologous to pesticidal-like nucleic acid molecules of the invention. BLAST protein searches can be performed with the BLASTX program, score=50, wordlength=3, to obtain amino acid sequences homologous to pesticidal protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25:3389. Alternatively, PSI-Blast can be used to perform an iterated search that detects distant relationships between molecules. See Altschul et al. (1997) supra. When utilizing BLAST, Gapped BLAST, and PSI-Blast programs, the default parameters of the respective programs (e.g., BLASTX and BLASTN) can be used. Alignment may also be performed manually by inspection.
[0026] Another non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the ClustalW algorithm (Higgins et al. (1994) Nucleic Acids Res. 22:4673-4680). ClustalW compares sequences and aligns the entirety of the amino acid or DNA sequence, and thus can provide data about the sequence conservation of the entire amino acid sequence. The ClustalW algorithm is used in several commercially available DNA/amino acid analysis software packages, such as the ALIGNX module of the Vector NTI Program Suite (Invitrogen Corporation, Carlsbad, Calif.). After alignment of amino acid sequences with ClustalW, the percent amino acid identity can be assessed. A non-limiting example of a software program useful for analysis of ClustalW alignments is GENEDOC®. GENEDOC® (Karl Nicholas) allows assessment of amino acid (or DNA) similarity and identity between multiple proteins. Another 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 into the ALIGN program (version 2.0), which is part of the GCG Wisconsin Genetics Software Package, Version 10 (available from Accelrys, Inc., 9685 Scranton Rd., San Diego, Calif., USA). 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.
[0027] Unless otherwise stated, GAP Version 10, which uses the algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48(3):443-453, will be used to determine sequence identity or similarity using the following parameters: % identity and % similarity for a nucleotide sequence using GAP Weight of 50 and Length Weight of 3, and the nwsgapdna.cmp scoring matrix; % identity or % similarity for an amino acid sequence using GAP weight of 8 and length weight of 2, and the BLOSUM62 scoring program. Equivalent programs may also be used. By "equivalent program" is intended any sequence comparison program that, for any two sequences in question, generates an alignment having identical nucleotide residue matches and an identical percent sequence identity when compared to the corresponding alignment generated by GAP Version 10. The invention also encompasses variant nucleic acid molecules. "Variants" of the pesticidal protein encoding nucleotide sequences include those sequences that encode the pesticidal proteins disclosed herein but that differ conservatively because of the degeneracy of the genetic code as well as those that are sufficiently identical as discussed above. Naturally occurring allelic variants can be identified with the use of well-known molecular biology techniques, such as polymerase chain reaction (PCR) and hybridization techniques as outlined below. Variant nucleotide sequences also include synthetically derived nucleotide sequences that have been generated, for example, by using site-directed mutagenesis but which still encode the pesticidal proteins disclosed in the present invention as discussed below. Variant proteins encompassed by the present invention are biologically active, that is they continue to possess the desired biological activity of the native protein, that is, retaining pesticidal activity. By "retains activity" is intended that the variant will have at least about 30%, at least about 50%, at least about 70%, or at least about 80% of the pesticidal activity of the native protein. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83: 2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety.
[0028] The skilled artisan will further appreciate that changes can be introduced by mutation of the nucleotide sequences of the invention thereby leading to changes in the amino acid sequence of the encoded pesticidal proteins, without altering the biological activity of the proteins. Thus, variant isolated nucleic acid molecules can be created by introducing one or more nucleotide substitutions, additions, or deletions into the corresponding nucleotide sequence disclosed herein, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Such variant nucleotide sequences are also encompassed by the present invention.
[0029] For example, conservative amino acid substitutions may be made at one or more, predicted, nonessential amino acid residues. A "nonessential" amino acid residue is a residue that can be altered from the wild-type sequence of a pesticidal protein without altering the biological activity, whereas an "essential" amino acid residue is required for biological activity. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).
[0030] Delta-endotoxins generally have five conserved sequence domains, and three conserved structural domains (see, for example, de Maagd et al. (2001) Trends Genetics 17:193-199). The first conserved structural domain consists of seven alpha helices and is involved in membrane insertion and pore formation. Domain II consists of three beta-sheets arranged in a Greek key configuration, and domain III consists of two antiparallel beta-sheets in "jelly-roll" formation (de Maagd et al., 2001, supra). Domains II and III are involved in receptor recognition and binding, and are therefore considered determinants of toxin specificity.
[0031] Amino acid substitutions may be made in nonconserved regions that retain function. In general, such substitutions would not be made for conserved amino acid residues, or for amino acid residues residing within a conserved motif, where such residues are essential for protein activity. Examples of residues that are conserved and that may be essential for protein activity include, for example, residues that are identical between all proteins contained in an alignment of similar or related toxins to the sequences of the invention (e.g., residues that are identical in an alignment of homologous proteins). Examples of residues that are conserved but that may allow conservative amino acid substitutions and still retain activity include, for example, residues that have only conservative substitutions between all proteins contained in an alignment of similar or related toxins to the sequences of the invention (e.g., residues that have only conservative substitutions between all proteins contained in the alignment homologous proteins). However, one of skill in the art would understand that functional variants may have minor conserved or nonconserved alterations in the conserved residues.
[0032] Alternatively, variant nucleotide sequences can be made by introducing mutations randomly along all or part of the coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for ability to confer pesticidal activity to identify mutants that retain activity. Following mutagenesis, the encoded protein can be expressed recombinantly, and the activity of the protein can be determined using standard assay techniques.
[0033] Using methods such as PCR, hybridization, and the like corresponding pesticidal sequences can be identified, isolated, or recovered from a sample (e.g., a sample containing nucleic acid sequences, such as a biological sample), such sequences having substantial identity to the sequences of the invention (e.g., at least about 70%, at least about 75%, 80%, 85%, 90%, 95% or more sequence identity across the entirety of the reference sequence) and having or conferring pesticidal activity. See, for example, Sambrook and Russell (2001) Molecular Cloning: A Laboratory Manual. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY) and Innis, et al. (1990) PCR Protocols: A Guide to Methods and Applications (Academic Press, NY).
[0034] In a hybridization method, all or part of the pesticidal nucleotide sequence can be used to screen cDNA or genomic libraries. Methods for construction of such cDNA and genomic libraries are generally known in the art and are disclosed in Sambrook and Russell, 2001, supra. The so-called hybridization probes may be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and may be labeled with a detectable group such as 32P, or any other detectable marker, such as other radioisotopes, a fluorescent compound, an enzyme, or an enzyme co-factor. Probes for hybridization can be made by labeling synthetic oligonucleotides based on the known pesticidal protein-encoding nucleotide sequence disclosed herein. Degenerate primers designed on the basis of conserved nucleotides or amino acid residues in the nucleotide sequence or encoded amino acid sequence can additionally be used. The probe typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, at least about 25, at least about 50, 75, 100, 125, 150, 175, or 200 consecutive nucleotides of nucleotide sequence encoding a pesticidal protein of the invention or a fragment or variant thereof. Methods for the preparation of probes for hybridization are generally known in the art and are disclosed in Sambrook and Russell, 2001, supra herein incorporated by reference.
[0035] For example, an entire pesticidal protein sequence disclosed herein, or one or more portions thereof, may be used as a probe capable of specifically hybridizing to corresponding pesticidal protein-like sequences and messenger RNAs. To achieve specific hybridization under a variety of conditions, such probes include sequences that are unique and are preferably at least about 10 nucleotides in length, or at least about 20 nucleotides in length. Such probes may be used to amplify corresponding pesticidal sequences from a chosen organism or sample by PCR. This technique may be used to isolate additional coding sequences from a desired organism or as a diagnostic assay to determine the presence of coding sequences in an organism. Hybridization techniques include hybridization screening of plated DNA libraries (either plaques or colonies; see, for example, Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).
[0036] Hybridization of such sequences may be carried out under stringent conditions. By "stringent conditions" or "stringent hybridization conditions" is intended conditions under which a probe will hybridize to its target sequence to a detectably greater degree than to other sequences (e.g., at least 2-fold over background). Stringent conditions are sequence-dependent and will be different in different circumstances. By controlling the stringency of the hybridization and/or washing conditions, target sequences that are 100% complementary to the probe can be identified (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity are detected (heterologous probing). Generally, a probe is less than about 1000 nucleotides in length, preferably less than 500 nucleotides in length.
[0037] Typically, stringent conditions will be those in which the salt concentration is less than about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes (e.g., 10 to 50 nucleotides) and at least about 60° C. for long probes (e.g., greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization with a buffer solution of 30 to 35% formamide, 1 M NaCl, 1% SDS (sodium dodecyl sulphate) at 37° C., and a wash in 1× to 2×SSC (20×SSC=3.0 M NaCl/0.3 M trisodium citrate) at 50 to 55° C. Exemplary moderate stringency conditions include hybridization in 40 to 45% formamide, 1.0 M NaCl, 1% SDS at 37° C., and a wash in 0.5× to 1×SSC at 55 to 60° C. Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60 to 65° C. Optionally, wash buffers may comprise about 0.1% to about 1% SDS. Duration of hybridization is generally less than about 24 hours, usually about 4 to about 12 hours.
[0038] Specificity is typically the function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution. For DNA-DNA hybrids, the Tm can be approximated from the equation of Meinkoth and Wahl (1984) Anal. Biochem. 138:267-284: Tm=81.5° C.+16.6 (log M)+0.41 (% GC)-0.61 (% form)-500/L; where M is the molarity of monovalent cations, % GC is the percentage of guanosine and cytosine nucleotides in the DNA, % form is the percentage of formamide in the hybridization solution, and L is the length of the hybrid in base pairs. The Tm is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. Tm is reduced by about 1° C. for each 1% of mismatching; thus, Tm, hybridization, and/or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with >90% identity are sought, the Tm can be decreased 10° C. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (Tm) for the specific sequence and its complement at a defined ionic strength and pH. However, severely stringent conditions can utilize a hybridization and/or wash at 1, 2, 3, or 4° C. lower than the thermal melting point (Tm); moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9, or 10° C. lower than the thermal melting point (Tm); low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 15, or 20° C. lower than the thermal melting point (Tm). Using the equation, hybridization and wash compositions, and desired Tm, those of ordinary skill will understand that variations in the stringency of hybridization and/or wash solutions are inherently described. If the desired degree of mismatching results in a Tm of less than 45° C. (aqueous solution) or 32° C. (formamide solution), it is preferred to increase the SSC concentration so that a higher temperature can be used. An extensive guide to the hybridization of nucleic acids is found in Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology--Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, New York); and Ausubel et al., eds. (1995) Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience, New York). See Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).
Isolated Proteins and Variants and Fragments Thereof
[0039] Pesticidal proteins are also encompassed within the present invention. By "pesticidal protein" is intended a protein having the amino acid sequence set forth in SEQ ID NO:28-62. Fragments, biologically active portions, and variants thereof are also provided, and may be used to practice the methods of the present invention. An "isolated protein" is used to refer to a protein that is no longer in its natural environment, for example in vitro or in a recombinant bacterial or plant host cell.
[0040] "Fragments" or "biologically active portions" include polypeptide fragments comprising amino acid sequences sufficiently identical to the amino acid sequence set forth in any of SEQ ID NO:28-62 and that exhibit pesticidal activity. A biologically active portion of a pesticidal protein can be a polypeptide that is, for example, 10, 25, 50, 100 or more amino acids in length. Such biologically active portions can be prepared by recombinant techniques and evaluated for pesticidal activity. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety. As used here, a fragment comprises at least 8 contiguous amino acids of SEQ ID NO:28-62. The invention encompasses other fragments, however, such as any fragment in the protein greater than about 10, 20, 30, 50, 100, 150, 200, 250, 300, 350, 400, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, or 1300 amino acids.
[0041] By "variants" is intended proteins or polypeptides having an amino acid sequence that is at least about 60%, 65%, about 70%, 75%, about 80%, 85%, about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of any of SEQ ID NO:28-62. Variants also include polypeptides encoded by a nucleic acid molecule that hybridizes to the nucleic acid molecule of SEQ ID NO:1-27, or a complement thereof, under stringent conditions. Variants include polypeptides that differ in amino acid sequence due to mutagenesis. Variant proteins encompassed by the present invention are biologically active, that is they continue to possess the desired biological activity of the native protein, that is, retaining pesticidal activity. In some embodiments, the variants have improved activity. Methods for measuring pesticidal activity are well known in the art. See, for example, Czapla and Lang (1990) J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J. 252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293; and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety.
[0042] Bacterial genes, such as the axmi genes of this invention, quite often possess multiple methionine initiation codons in proximity to the start of the open reading frame. Often, translation initiation at one or more of these start codons will lead to generation of a functional protein. These start codons can include ATG codons. However, bacteria such as Bacillus sp. also recognize the codon GTG as a start codon, and proteins that initiate translation at GTG codons contain a methionine at the first amino acid. On rare occasions, translation in bacterial systems can initiate at a TTG codon, though in this event the TTG encodes a methionine. Furthermore, it is not often determined a priori which of these codons are used naturally in the bacterium. Thus, it is understood that use of one of the alternate methionine codons may also lead to generation of pesticidal proteins. These pesticidal proteins are encompassed in the present invention and may be used in the methods of the present invention. It will be understood that, when expressed in plants, it will be necessary to alter the alternate start codon to ATG for proper translation.
[0043] Antibodies to the polypeptides of the present invention, or to variants or fragments thereof, are also encompassed. Methods for producing antibodies are well known in the art (see, for example, Harlow and Lane (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.; U.S. Pat. No. 4,196,265).
Altered or Improved Variants
[0044] It is recognized that DNA sequences of a pesticidal protein may be altered by various methods, and that these alterations may result in DNA sequences encoding proteins with amino acid sequences different than that encoded by a pesticidal protein of the present invention. This protein may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions of one or more amino acids of SEQ ID NO:28-62, including up to about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 100, about 105, about 110, about 115, about 120, about 125, about 130 or more amino acid substitutions, deletions or insertions.
[0045] Methods for such manipulations are generally known in the art. For example, amino acid sequence variants of a pesticidal protein can be prepared by mutations in the DNA. This may also be accomplished by one of several forms of mutagenesis and/or in directed evolution. In some aspects, the changes encoded in the amino acid sequence will not substantially affect the function of the protein. Such variants will possess the desired pesticidal activity. However, it is understood that the ability of a pesticidal protein to confer pesticidal activity may be improved by the use of such techniques upon the compositions of this invention. For example, one may express a pesticidal protein in host cells that exhibit high rates of base misincorporation during DNA replication, such as XL-1 Red (Stratagene). After propagation in such strains, one can isolate the toxin DNA (for example by preparing plasmid DNA, or by amplifying by PCR and cloning the resulting PCR fragment into a vector), culture the toxin mutations in a non-mutagenic strain, and identify mutated toxin genes with pesticidal activity, for example by performing an assay to test for pesticidal activity. Generally, the protein is mixed and used in feeding assays. See, for example Marrone et al. (1985) J. of Economic Entomology 78:290-293. Such assays can include contacting plants with one or more pests and determining the plant's ability to survive and/or cause the death of the pests. Examples of mutations that result in increased toxicity are found in Schnepf et al. (1998) Microbiol. Mol. Biol. Rev. 62:775-806.
[0046] Alternatively, alterations may be made to the protein sequence of many proteins at the amino or carboxy terminus without substantially affecting activity. This can include insertions, deletions, or alterations introduced by modern molecular methods, such as PCR, including PCR amplifications that alter or extend the protein coding sequence by virtue of inclusion of amino acid encoding sequences in the oligonucleotides utilized in the PCR amplification. Alternatively, the protein sequences added can include entire protein-coding sequences, such as those used commonly in the art to generate protein fusions. Such fusion proteins are often used to (1) increase expression of a protein of interest (2) introduce a binding domain, enzymatic activity, or epitope to facilitate either protein purification, protein detection, or other experimental uses known in the art (3) target secretion or translation of a protein to a subcellular organelle, such as the periplasmic space of Gram-negative bacteria, or the endoplasmic reticulum of eukaryotic cells, the latter of which often results in glycosylation of the protein.
[0047] Variant nucleotide and amino acid sequences of the present invention also encompass sequences derived from mutagenic and recombinogenic procedures such as DNA shuffling. With such a procedure, one or more different pesticidal protein coding regions can be used to create a new pesticidal protein possessing the desired properties. In this manner, libraries of recombinant polynucleotides are generated from a population of related sequence polynucleotides comprising sequence regions that have substantial sequence identity and can be homologously recombined in vitro or in vivo. For example, using this approach, sequence motifs encoding a domain of interest may be shuffled between a pesticidal gene of the invention and other known pesticidal genes to obtain a new gene coding for a protein with an improved property of interest, such as an increased insecticidal activity. Strategies for such DNA shuffling are known in the art. See, for example, Stemmer (1994) Proc. Natl. Acad. Sci. USA 91:10747-10751; Stemmer (1994) Nature 370:389-391; Crameri et al. (1997) Nature Biotech. 15:436-438; Moore et al. (1997) J. Mol. Biol. 272:336-347; Zhang et al. (1997) Proc. Natl. Acad. Sci. USA 94:4504-4509; Crameri et al. (1998) Nature 391:288-291; and U.S. Pat. Nos. 5,605,793 and 5,837,458.
[0048] Domain swapping or shuffling is another mechanism for generating altered delta-endotoxin proteins. Domains may be swapped between delta-endotoxin proteins, resulting in hybrid or chimeric toxins with improved pesticidal activity or target spectrum. Methods for generating recombinant proteins and testing them for pesticidal activity are well known in the art (see, for example, Naimov et al. (2001) Appl. Environ. Microbiol. 67:5328-5330; de Maagd et al. (1996) Appl. Environ. Microbiol. 62:1537-1543; Ge et al. (1991) J. Biol. Chem. 266:17954-17958; Schnepf et al. (1990) J. Biol. Chem. 265:20923-20930; Rang et al. 91999) Appl. Environ. Microbiol. 65:2918-2925).
[0049] In yet another embodiment, variant nucleotide and/or amino acid sequences can be obtained using one or more of error-prone PCR, oligonucleotide-directed mutagenesis, assembly PCR, sexual PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive ensemble mutagenesis, exponential ensemble mutagenesis, site-specific mutagenesis, gene reassembly, gene site saturation mutagenesis, permutational mutagenesis, synthetic ligation reassembly (SLR), recombination, recursive sequence recombination, phosphothioate-modified DNA mutagenesis, uracil-containing template mutagenesis, gapped duplex mutagenesis, point mismatch repair mutagenesis, repair-deficient host strain mutagenesis, chemical mutagenesis, radiogenic mutagenesis, deletion mutagenesis, restriction-selection mutagenesis, restriction-purification mutagenesis, artificial gene synthesis, ensemble mutagenesis, chimeric nucleic acid multimer creation, and the like.
Vectors
[0050] A pesticidal sequence of the invention may be provided in an expression cassette for expression in a plant of interest. By "plant expression cassette" is intended a DNA construct that is capable of resulting in the expression of a protein from an open reading frame in a plant cell. Typically these contain a promoter and a coding sequence. Often, such constructs will also contain a 3' untranslated region. Such constructs may contain a "signal sequence" or "leader sequence" to facilitate co-translational or post-translational transport of the peptide to certain intracellular structures such as the chloroplast (or other plastid), endoplasmic reticulum, or Golgi apparatus.
[0051] By "signal sequence" is intended a sequence that is known or suspected to result in cotranslational or post-translational peptide transport across the cell membrane. In eukaryotes, this typically involves secretion into the Golgi apparatus, with some resulting glycosylation. Insecticidal toxins of bacteria are often synthesized as protoxins, which are protolytically activated in the gut of the target pest (Chang (1987) Methods Enzymol. 153:507-516). In some embodiments of the present invention, the signal sequence is located in the native sequence, or may be derived from a sequence of the invention. By "leader sequence" is intended any sequence that when translated, results in an amino acid sequence sufficient to trigger co-translational transport of the peptide chain to a sub-cellular organelle. Thus, this includes leader sequences targeting transport and/or glycosylation by passage into the endoplasmic reticulum, passage to vacuoles, plastids including chloroplasts, mitochondria, and the like.
[0052] By "plant transformation vector" is intended a DNA molecule that is necessary for efficient transformation of a plant cell. Such a molecule may consist of one or more plant expression cassettes, and may be organized into more than one "vector" DNA molecule. For example, binary vectors are plant transformation vectors that utilize two non-contiguous DNA vectors to encode all requisite cis- and trans-acting functions for transformation of plant cells (Hellens and Mullineaux (2000) Trends in Plant Science 5:446-451). "Vector" refers to a nucleic acid construct designed for transfer between different host cells. "Expression vector" refers to a vector that has the ability to incorporate, integrate and express heterologous DNA sequences or fragments in a foreign cell. The cassette will include 5' and 3' regulatory sequences operably linked to a sequence of the invention. By "operably linked" is intended a functional linkage between a promoter and a second sequence, wherein the promoter sequence initiates and mediates transcription of the DNA sequence corresponding to the second sequence. Generally, operably linked means that the nucleic acid sequences being linked are contiguous and, where necessary to join two protein coding regions, contiguous and in the same reading frame. The cassette may additionally contain at least one additional gene to be cotransformed into the organism. Alternatively, the additional gene(s) can be provided on multiple expression cassettes.
[0053] "Promoter" refers to a nucleic acid sequence that functions to direct transcription of a downstream coding sequence. The promoter together with other transcriptional and translational regulatory nucleic acid sequences (also termed "control sequences") are necessary for the expression of a DNA sequence of interest.
[0054] Such an expression cassette is provided with a plurality of restriction sites for insertion of the pesticidal sequence to be under the transcriptional regulation of the regulatory regions.
[0055] The expression cassette will include in the 5'-3' direction of transcription, a transcriptional and translational initiation region (i.e., a promoter), a DNA sequence of the invention, and a translational and transcriptional termination region (i.e., termination region) functional in plants. The promoter may be native or analogous, or foreign or heterologous, to the plant host and/or to the DNA sequence of the invention. Additionally, the promoter may be the natural sequence or alternatively a synthetic sequence. Where the promoter is "native" or "homologous" to the plant host, it is intended that the promoter is found in the native plant into which the promoter is introduced. Where the promoter is "foreign" or "heterologous" to the DNA sequence of the invention, it is intended that the promoter is not the native or naturally occurring promoter for the operably linked DNA sequence of the invention.
[0056] The termination region may be native with the transcriptional initiation region, may be native with the operably linked DNA sequence of interest, may be native with the plant host, or may be derived from another source (i.e., foreign or heterologous to the promoter, the DNA sequence of interest, the plant host, or any combination thereof). Convenient termination regions are available from the Ti-plasmid of A. tumefaciens, such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al. (1991) Mol. Gen. Genet. 262:141-144; Proudfoot (1991) Cell 64:671-674; Sanfacon et al. (1991) Genes Dev. 5:141-149; Mogen et al. (1990) Plant Cell 2:1261-1272; Munroe et al. (1990) Gene 91:151-158; Ballas et al. (1989) Nucleic Acids Res. 17:7891-7903; and Joshi et al. (1987) Nucleic Acid Res. 15:9627-9639.
[0057] Where appropriate, the gene(s) may be optimized for increased expression in the transformed host cell. That is, the genes can be synthesized using host cell-preferred codons for improved expression, or may be synthesized using codons at a host-preferred codon usage frequency. Generally, the GC content of the gene will be increased. See, for example, Campbell and Gowri (1990) Plant Physiol. 92:1-11 for a discussion of host-preferred codon usage. Methods are available in the art for synthesizing plant-preferred genes. See, for example, U.S. Pat. Nos. 5,380,831, and 5,436,391, and Murray et al. (1989) Nucleic Acids Res. 17:477-498, herein incorporated by reference.
[0058] In one embodiment, the pesticidal sequence is targeted to the chloroplast for expression. In this manner, where the pesticidal sequence is not directly inserted into the chloroplast, the expression cassette will additionally contain a nucleic acid encoding a transit peptide to direct the pesticidal sequence to the chloroplasts. Such transit peptides are known in the art. See, for example, Von Heijne et al. (1991) Plant Mol. Biol. Rep. 9:104-126; Clark et al. (1989) J. Biol. Chem. 264:17544-17550; Della-Cioppa et al. (1987) Plant Physiol. 84:965-968; Romer et al. (1993) Biochem. Biophys. Res. Commun. 196:1414-1421; and Shah et al. (1986) Science 233:478-481.
[0059] The pesticidal gene to be targeted to the chloroplast may be optimized for expression in the chloroplast to account for differences in codon usage between the plant nucleus and this organelle. In this manner, the nucleic acids of interest may be synthesized using chloroplast-preferred codons. See, for example, U.S. Pat. No. 5,380,831, herein incorporated by reference.
Plant Transformation
[0060] Methods of the invention involve introducing a nucleotide construct into a plant. By "introducing" is intended to present to the plant the nucleotide construct in such a manner that the construct gains access to the interior of a cell of the plant. The methods of the invention do not require that a particular method for introducing a nucleotide construct to a plant is used, only that the nucleotide construct gains access to the interior of at least one cell of the plant. Methods for introducing nucleotide constructs into plants are known in the art including, but not limited to, stable transformation methods, transient transformation methods, and virus-mediated methods.
[0061] By "plant" is intended whole plants, plant organs (e.g., leaves, stems, roots, etc.), seeds, plant cells, propagules, embryos and progeny of the same. Plant cells can be differentiated or undifferentiated (e.g. callus, suspension culture cells, protoplasts, leaf cells, root cells, phloem cells, pollen).
[0062] "Transgenic plants" or "transformed plants" or "stably transformed" plants or cells or tissues refers to plants that have incorporated or integrated exogenous nucleic acid sequences or DNA fragments into the plant cell. These nucleic acid sequences include those that are exogenous, or not present in the untransformed plant cell, as well as those that may be endogenous, or present in the untransformed plant cell. "Heterologous" generally refers to the nucleic acid sequences that are not endogenous to the cell or part of the native genome in which they are present, and have been added to the cell by infection, transfection, microinjection, electroporation, microprojection, or the like.
[0063] The transgenic plants of the invention express one or more of the pesticidal sequences disclosed herein. In various embodiments, the transgenic plant further comprises one or more additional genes for insect resistance, for example, one or more additional genes for controlling coleopteran, lepidopteran, heteropteran, or nematode pests. It will be understood by one of skill in the art that the transgenic plant may comprise any gene imparting an agronomic trait of interest.
[0064] Transformation of plant cells can be accomplished by one of several techniques known in the art. The pesticidal gene of the invention may be modified to obtain or enhance expression in plant cells. Typically a construct that expresses such a protein would contain a promoter to drive transcription of the gene, as well as a 3' untranslated region to allow transcription termination and polyadenylation. The organization of such constructs is well known in the art. In some instances, it may be useful to engineer the gene such that the resulting peptide is secreted, or otherwise targeted within the plant cell. For example, the gene can be engineered to contain a signal peptide to facilitate transfer of the peptide to the endoplasmic reticulum. It may also be preferable to engineer the plant expression cassette to contain an intron, such that mRNA processing of the intron is required for expression.
[0065] Typically this "plant expression cassette" will be inserted into a "plant transformation vector". This plant transformation vector may be comprised of one or more DNA vectors needed for achieving plant transformation. For example, it is a common practice in the art to utilize plant transformation vectors that are comprised of more than one contiguous DNA segment. These vectors are often referred to in the art as "binary vectors". Binary vectors as well as vectors with helper plasmids are most often used for Agrobacterium-mediated transformation, where the size and complexity of DNA segments needed to achieve efficient transformation is quite large, and it is advantageous to separate functions onto separate DNA molecules. Binary vectors typically contain a plasmid vector that contains the cis-acting sequences required for T-DNA transfer (such as left border and right border), a selectable marker that is engineered to be capable of expression in a plant cell, and a "gene of interest" (a gene engineered to be capable of expression in a plant cell for which generation of transgenic plants is desired). Also present on this plasmid vector are sequences required for bacterial replication. The cis-acting sequences are arranged in a fashion to allow efficient transfer into plant cells and expression therein. For example, the selectable marker gene and the pesticidal sequence are located between the left and right borders. Often a second plasmid vector contains the trans-acting factors that mediate T-DNA transfer from Agrobacterium to plant cells. This plasmid often contains the virulence functions (Vir genes) that allow infection of plant cells by Agrobacterium, and transfer of DNA by cleavage at border sequences and vir-mediated DNA transfer, as is understood in the art (Hellens and Mullineaux (2000) Trends in Plant Science 5:446-451). Several types of Agrobacterium strains (e.g. LBA4404, GV3101, EHA101, EHA105, etc.) can be used for plant transformation. The second plasmid vector is not necessary for transforming the plants by other methods such as microprojection, microinjection, electroporation, polyethylene glycol, etc.
[0066] In general, plant transformation methods involve transferring heterologous DNA into target plant cells (e.g. immature or mature embryos, suspension cultures, undifferentiated callus, protoplasts, etc.), followed by applying a maximum threshold level of appropriate selection (depending on the selectable marker gene) to recover the transformed plant cells from a group of untransformed cell mass. Explants are typically transferred to a fresh supply of the same medium and cultured routinely. Subsequently, the transformed cells are differentiated into shoots after placing on regeneration medium supplemented with a maximum threshold level of selecting agent. The shoots are then transferred to a selective rooting medium for recovering rooted shoot or plantlet. The transgenic plantlet then grows into a mature plant and produces fertile seeds (e.g. Hiei et al. (1994) The Plant Journal 6:271-282; Ishida et al. (1996) Nature Biotechnology 14:745-750). Explants are typically transferred to a fresh supply of the same medium and cultured routinely. A general description of the techniques and methods for generating transgenic plants are found in Ayres and Park (1994) Critical Reviews in Plant Science 13:219-239 and Bommineni and Jauhar (1997) Maydica 42:107-120. Since the transformed material contains many cells; both transformed and non-transformed cells are present in any piece of subjected target callus or tissue or group of cells. The ability to kill non-transformed cells and allow transformed cells to proliferate results in transformed plant cultures. Often, the ability to remove non-transformed cells is a limitation to rapid recovery of transformed plant cells and successful generation of transgenic plants.
[0067] Transformation protocols as well as protocols for introducing nucleotide sequences into plants may vary depending on the type of plant or plant cell, i.e., monocot or dicot, targeted for transformation. Generation of transgenic plants may be performed by one of several methods, including, but not limited to, microinjection, electroporation, direct gene transfer, introduction of heterologous DNA by Agrobacterium into plant cells (Agrobacterium-mediated transformation), bombardment of plant cells with heterologous foreign DNA adhered to particles, ballistic particle acceleration, aerosol beam transformation (U.S. Published Application No. 20010026941; U.S. Pat. No. 4,945,050; International Publication No. WO 91/00915; U.S. Published Application No. 2002015066), Lec1 transformation, and various other non-particle direct-mediated methods to transfer DNA.
[0068] Methods for transformation of chloroplasts are known in the art. See, for example, Svab et al. (1990) Proc. Natl. Acad. Sci. USA 87:8526-8530; Svab and Maliga (1993) Proc. Natl. Acad. Sci. USA 90:913-917; Svab and Maliga (1993) EMBO J. 12:601-606. The method relies on particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination. Additionally, plastid transformation can be accomplished by transactivation of a silent plastid-borne transgene by tissue-preferred expression of a nuclear-encoded and plastid-directed RNA polymerase. Such a system has been reported in McBride et al. (1994) Proc. Natl. Acad. Sci. USA 91:7301-7305.
[0069] Following integration of heterologous foreign DNA into plant cells, one then applies a maximum threshold level of appropriate selection in the medium to kill the untransformed cells and separate and proliferate the putatively transformed cells that survive from this selection treatment by transferring regularly to a fresh medium. By continuous passage and challenge with appropriate selection, one identifies and proliferates the cells that are transformed with the plasmid vector. Molecular and biochemical methods can then be used to confirm the presence of the integrated heterologous gene of interest into the genome of the transgenic plant.
[0070] The cells that have been transformed may be grown into plants in accordance with conventional ways. See, for example, McCormick et al. (1986) Plant Cell Reports 5:81-84. These plants may then be grown, and either pollinated with the same transformed strain or different strains, and the resulting hybrid having constitutive expression of the desired phenotypic characteristic identified. Two or more generations may be grown to ensure that expression of the desired phenotypic characteristic is stably maintained and inherited and then seeds harvested to ensure expression of the desired phenotypic characteristic has been achieved. In this manner, the present invention provides transformed seed (also referred to as "transgenic seed") having a nucleotide construct of the invention, for example, an expression cassette of the invention, stably incorporated into their genome.
Evaluation of Plant Transformation
[0071] Following introduction of heterologous foreign DNA into plant cells, the transformation or integration of heterologous gene in the plant genome is confirmed by various methods such as analysis of nucleic acids, proteins and metabolites associated with the integrated gene.
[0072] PCR analysis is a rapid method to screen transformed cells, tissue or shoots for the presence of incorporated gene at the earlier stage before transplanting into the soil (Sambrook and Russell (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.). PCR is carried out using oligonucleotide primers specific to the gene of interest or Agrobacterium vector background, etc.
[0073] Plant transformation may be confirmed by Southern blot analysis of genomic DNA (Sambrook and Russell, 2001, supra). In general, total DNA is extracted from the transformant, digested with appropriate restriction enzymes, fractionated in an agarose gel and transferred to a nitrocellulose or nylon membrane. The membrane or "blot" is then probed with, for example, radiolabeled 32P target DNA fragment to confirm the integration of introduced gene into the plant genome according to standard techniques (Sambrook and Russell, 2001, supra).
[0074] In Northern blot analysis, RNA is isolated from specific tissues of transformant, fractionated in a formaldehyde agarose gel, and blotted onto a nylon filter according to standard procedures that are routinely used in the art (Sambrook and Russell, 2001, supra). Expression of RNA encoded by the pesticidal sequence is then tested by hybridizing the filter to a radioactive probe derived from a toxin, by methods known in the art (Sambrook and Russell, 2001, supra).
[0075] Western blot, biochemical assays and the like may be carried out on the transgenic plants to confirm the presence of protein encoded by the pesticidal gene by standard procedures (Sambrook and Russell, 2001, supra) using antibodies that bind to one or more epitopes present on the pesticidal protein.
Pesticidal Activity in Plants
[0076] In another aspect of the invention, one may generate transgenic plants expressing a toxin that has pesticidal activity. Methods described above by way of example may be utilized to generate transgenic plants, but the manner in which the transgenic plant cells are generated is not critical to this invention. Methods known or described in the art such as Agrobacterium-mediated transformation, biolistic transformation, and non-particle-mediated methods may be used at the discretion of the experimenter. Plants expressing a pesticidal sequence may be isolated by common methods described in the art, for example by transformation of callus, selection of transformed callus, and regeneration of fertile plants from such transgenic callus. In such process, one may use any gene as a selectable marker so long as its expression in plant cells confers ability to identify or select for transformed cells.
[0077] A number of markers have been developed for use with plant cells, such as resistance to chloramphenicol, the aminoglycoside G418, hygromycin, or the like. Other genes that encode a product involved in chloroplast metabolism may also be used as selectable markers. For example, genes that provide resistance to plant herbicides such as glyphosate, bromoxynil, or imidazolinone may find particular use. Such genes have been reported (Stalker et al. (1985) J. Biol. Chem. 263:6310-6314 (bromoxynil resistance nitrilase gene); and Sathasivan et al. (1990) Nucl. Acids Res. 18:2188 (AHAS imidazolinone resistance gene). Additionally, the genes disclosed herein are useful as markers to assess transformation of bacterial or plant cells. Methods for detecting the presence of a transgene in a plant, plant organ (e.g., leaves, stems, roots, etc.), seed, plant cell, propagule, embryo or progeny of the same are well known in the art. In one embodiment, the presence of the transgene is detected by testing for pesticidal activity.
[0078] Fertile plants expressing a pesticidal sequence may be tested for pesticidal activity, and the plants showing optimal activity selected for further breeding. Methods are available in the art to assay for pest activity. Generally, the protein is mixed and used in feeding assays. See, for example Marrone et al. (1985) J. of Economic Entomology 78:290-293.
[0079] The present invention may be used for transformation of any plant species, including, but not limited to, monocots and dicots. Examples of plants of interest include, but are not limited to, corn (maize), sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, and oilseed rape, Brassica sp., alfalfa, rye, millet, safflower, peanuts, sweet potato, cassaya, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana, avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond, oats, vegetables, ornamentals, and conifers.
[0080] Vegetables include, but are not limited to, tomatoes, lettuce, green beans, lima beans, peas, and members of the genus Curcumis such as cucumber, cantaloupe, and musk melon. Ornamentals include, but are not limited to, azalea, hydrangea, hibiscus, roses, tulips, daffodils, petunias, carnation, poinsettia, and chrysanthemum. Preferably, plants of the present invention are crop plants (for example, maize, sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, oilseed rape., etc.).
Use in Pesticidal Control
[0081] General methods for employing strains comprising a nucleotide sequence of the present invention, or a variant thereof, in pesticide control or in engineering other organisms as pesticidal agents are known in the art. See, for example U.S. Pat. No. 5,039,523 and EP 0480762A2.
[0082] The Bacillus strains containing a nucleotide sequence of the present invention, or a variant thereof, or the microorganisms that have been genetically altered to contain a pesticidal gene and protein may be used for protecting agricultural crops and products from pests. In one aspect of the invention, whole, i.e., unlysed, cells of a toxin (pesticide)-producing organism are treated with reagents that prolong the activity of the pesticidal protein produced in the cell when the cell is applied to the environment of target pest(s).
[0083] Alternatively, the pesticide is produced by introducing a pesticidal gene into a cellular host. Expression of the pesticidal gene results, directly or indirectly, in the intracellular production and maintenance of the pesticide. In one aspect of this invention, these cells are then treated under conditions that prolong the activity of the pesticidal produced in the cell when the cell is applied to the environment of target pest(s). The resulting product retains the toxicity of the pesticidal protein. These naturally encapsulated pesticides may then be formulated in accordance with conventional techniques for application to the environment hosting a target pest, e.g., soil, water, and foliage of plants. See, for example EPA 0192319, and the references cited therein. Alternatively, one may formulate the cells expressing a gene of this invention such as to allow application of the resulting material as a pesticide.
[0084] Pesticidal Compositions
[0085] The active ingredients of the present invention are normally applied in the form of compositions and can be applied to the crop area or plant to be treated, simultaneously or in succession, with other compounds. These compounds can be fertilizers, weed killers, cryoprotectants, surfactants, detergents, pesticidal soaps, dormant oils, polymers, and/or time-release or biodegradable carrier formulations that permit long-term dosing of a target area following a single application of the formulation. They can also be selective herbicides, chemical insecticides, virucides, microbicides, amoebicides, pesticides, fungicides, bacteriocides, nematocides, molluscicides or mixtures of several of these preparations, if desired, together with further agriculturally acceptable carriers, surfactants or application-promoting adjuvants customarily employed in the art of formulation. Suitable carriers and adjuvants can be solid or liquid and correspond to the substances ordinarily employed in formulation technology, e.g. natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, binders or fertilizers Likewise the formulations may be prepared into edible "baits" or fashioned into pest "traps" to permit feeding or ingestion by a target pest of the pesticidal formulation.
[0086] Methods of applying an active ingredient of the present invention or an agrochemical composition of the present invention that contains at least one of the pesticidal proteins produced by the bacterial strains of the present invention include leaf application, seed coating and soil application. The number of applications and the rate of application depend on the intensity of infestation by the corresponding pest.
[0087] The composition may be formulated as a powder, dust, pellet, granule, spray, emulsion, colloid, solution, or such like, and may be prepared by such conventional means as desiccation, lyophilization, homogenation, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of cells comprising the polypeptide. In all such compositions that contain at least one such pesticidal polypeptide, the polypeptide may be present in a concentration of from about 1% to about 99% by weight.
[0088] Lepidopteran, dipteran, heteropteran, coleopteran, or nematode pests may be killed or reduced in numbers in a given area by the methods of the invention, or may be prophylactically applied to an environmental area to prevent infestation by a susceptible pest. Preferably the pest ingests, or is contacted with, a pesticidally-effective amount of the polypeptide. By "pesticidally-effective amount" is intended an amount of the pesticide that is able to bring about death to at least one pest, or to noticeably reduce pest growth, feeding, or normal physiological development. This amount will vary depending on such factors as, for example, the specific target pests to be controlled, the specific environment, location, plant, crop, or agricultural site to be treated, the environmental conditions, and the method, rate, concentration, stability, and quantity of application of the pesticidally-effective polypeptide composition. The formulations may also vary with respect to climatic conditions, environmental considerations, and/or frequency of application and/or severity of pest infestation.
[0089] The pesticide compositions described may be made by formulating either the bacterial cell, crystal and/or spore suspension, or isolated protein component with the desired agriculturally-acceptable carrier. The compositions may be formulated prior to administration in an appropriate means such as lyophilized, freeze-dried, desiccated, or in an aqueous carrier, medium or suitable diluent, such as saline or other buffer. The formulated compositions may be in the form of a dust or granular material, or a suspension in oil (vegetable or mineral), or water or oil/water emulsions, or as a wettable powder, or in combination with any other carrier material suitable for agricultural application. Suitable agricultural carriers can be solid or liquid and are well known in the art. The term "agriculturally-acceptable carrier" covers all adjuvants, inert components, dispersants, surfactants, tackifiers, binders, etc. that are ordinarily used in pesticide formulation technology; these are well known to those skilled in pesticide formulation. The formulations may be mixed with one or more solid or liquid adjuvants and prepared by various means, e.g., by homogeneously mixing, blending and/or grinding the pesticidal composition with suitable adjuvants using conventional formulation techniques. Suitable formulations and application methods are described in U.S. Pat. No. 6,468,523, herein incorporated by reference.
[0090] The plants can also be treated with one or more chemical compositions, including one or more herbicide, insecticides, or fungicides. Exemplary chemical compositions include: Fruits/Vegetables Herbicides: Atrazine, Bromacil, Diuron, Glyphosate, Linuron, Metribuzin, Simazine, Trifluralin, Fluazifop, Glufosinate, Halo sulfuron Gowan, Paraquat, Propyzamide, Sethoxydim, Butafenacil, Halosulfuron, Indaziflam; Fruits/Vegetables Insecticides: Aldicarb, Bacillus thuriengiensis, Carbaryl, Carbofuran, Chlorpyrifos, Cypermethrin, Deltamethrin, Diazinon, Malathion, Abamectin, Cyfluthrin/beta-cyfluthrin, Esfenvalerate, Lambda-cyhalothrin, Acequinocyl, Bifenazate, Methoxyfenozide, Novaluron, Chromafenozide, Thiacloprid, Dinotefuran, Fluacrypyrim, Tolfenpyrad, Clothianidin, Spirodiclofen, Gamma-cyhalothrin, Spiromesifen, Spinosad, Rynaxypyr, Cyazypyr, Spinoteram, Triflumuron, Spirotetramat, Imidacloprid, Flubendiamide, Thiodicarb, Metaflumizone, Sulfoxaflor, Cyflumetofen, Cyanopyrafen, Imidacloprid, Clothianidin, Thiamethoxam, Spinotoram, Thiodicarb, Flonicamid, Methiocarb, Emamectin-benzoate, Indoxacarb, Forthiazate, Fenamiphos, Cadusaphos, Pyriproxifen, Fenbutatin-oxid, Hexthiazox, Methomyl, 4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on; Fruits/Vegetables Fungicides: Carbendazim, Chlorothalonil, EBDCs, Sulphur, Thiophanate-methyl, Azoxystrobin, Cymoxanil, Fluazinam, Fosetyl, Iprodione, Kresoxim-methyl, Metalaxyl/mefenoxam, Trifloxystrobin, Ethaboxam, Iprovalicarb, Trifloxystrobin, Fenhexamid, Oxpoconazole fumarate, Cyazofamid, Fenamidone, Zoxamide, Picoxystrobin, Pyraclostrobin, Cyflufenamid, Boscalid; Cereals Herbicides: Isoproturon, Bromoxynil, Ioxynil, Phenoxies, Chlorsulfuron, Clodinafop, Diclofop, Diflufenican, Fenoxaprop, Florasulam, Fluoroxypyr, Metsulfuron, Triasulfuron, Flucarbazone, Iodosulfuron, Propoxycarbazone, Picolinafen, Mesosulfuron, Beflubutamid, Pinoxaden, Amidosulfuron, Thifensulfuron, Tribenuron, Flupyrsulfuron, Sulfosulfuron, Pyrasulfotole, Pyroxsulam, Flufenacet, Tralkoxydim, Pyroxasulfon; Cereals Fungicides: Carbendazim, Chlorothalonil, Azoxystrobin, Cyproconazole, Cyprodinil, Fenpropimorph, Epoxiconazole, Kresoxim-methyl, Quinoxyfen, Tebuconazole, Trifloxystrobin, Simeconazole, Picoxystrobin, Pyraclostrobin, Dimoxystrobin, Prothioconazole, Fluoxastrobin; Cereals Insecticides: Dimethoate, Lambda-cyhalthrin, Deltamethrin, alpha-Cypermethrin, B-cyfluthrin, Bifenthrin, Imidacloprid, Clothianidin, Thiamethoxam, Thiacloprid, Acetamiprid, Dinetofuran, Clorphyriphos, Metamidophos, Oxidemethon-methyl, Pirimicarb, Methiocarb; Maize Herbicides: Atrazine, Alachlor, Bromoxynil, Acetochlor, Dicamba, Clopyralid, (S-)Dimethenamid, Glufosinate, Glyphosate, Isoxaflutole, (S-)Metolachlor, Mesotrione, Nicosulfuron, Primisulfuron, Rimsulfuron, Sulcotrione, Foramsulfuron, Topramezone, Tembotrione, Saflufenacil, Thiencarbazone, Flufenacet, Pyroxasulfon; Maize Insecticides: Carbofuran, Chlorpyrifos, Bifenthrin, Cyazypyr, Fipronil, Imidacloprid, Lambda-Cyhalothrin, Tefluthrin, Terbufos, Thiamethoxam, Clothianidin, Spiromesifen, Flubendiamide, Triflumuron, Rynaxypyr, Deltamethrin, Thiodicarb, B-Cyfluthrin, Cypermethrin, Bifenthrin, Lufenuron, Triflumoron, Tefluthrin, Tebupirimphos, Ethiprole, Cyazypyr, Thiacloprid, Acetamiprid, Dinetofuran, Avermectin, Methiocarb, Spirodiclofen, Spirotetramat; Maize Fungicides: Fenitropan, Thiram, Prothioconazole, Tebuconazole, Trifloxystrobin; Rice Herbicides: Butachlor, Propanil, Azimsulfuron, Bensulfuron, Cyhalofop, Daimuron, Fentrazamide, Imazosulfuron, Mefenacet, Oxaziclomefone, Pyrazosulfuron, Pyributicarb, Quinclorac, Thiobencarb, Indanofan, Flufenacet, Fentrazamide, Halosulfuron, Oxaziclomefone, Benzobicyclon, Pyriftalid, Penoxsulam, Bispyribac, Oxadiargyl, Ethoxysulfuron, Pretilachlor, Mesotrione, Tefuryltrione, Oxadiazone, Fenoxaprop, Pyrimisulfan; Rice Insecticides: Diazinon, Fenitrothion, Fenobucarb, Monocrotophos, Benfuracarb, Buprofezin, Dinotefuran, Fipronil, Imidacloprid, Isoprocarb, Thiacloprid, Chromafenozide, Thiacloprid, Dinotefuran, Clothianidin, Ethiprole, Flubendiamide, Rynaxypyr, Deltamethrin, Acetamiprid, Thiamethoxam, Cyazypyr, Spinosad, Spinotoram, Emamectin-Benzoate, Cypermethrin, Chlorpyriphos, Cartap, Methamidophos, Etofenprox, Triazophos, 4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on, Carbofuran, Benfuracarb; Rice Fungicides: Thiophanate-methyl, Azoxystrobin, Carpropamid, Edifenphos, Ferimzone, Iprobenfos, Isoprothiolane, Pencycuron, Probenazole, Pyroquilon, Tricyclazole, Trifloxystrobin, Diclocymet, Fenoxanil, Simeconazole, Tiadinil; Cotton Herbicides: Diuron, Fluometuron, MSMA, Oxyfluorfen, Prometryn, Trifluralin, Carfentrazone, Clethodim, Fluazifop-butyl, Glyphosate, Norflurazon, Pendimethalin, Pyrithiobac-sodium, Trifloxysulfuron, Tepraloxydim, Glufosinate, Flumioxazin, Thidiazuron; Cotton Insecticides: Acephate, Aldicarb, Chlorpyrifos, Cypermethrin, Deltamethrin, Malathion, Monocrotophos, Abamectin, Acetamiprid, Emamectin Benzoate, Imidacloprid, Indoxacarb, Lambda-Cyhalothrin, Spinosad, Thiodicarb, Gamma-Cyhalothrin, Spiromesifen, Pyridalyl, Flonicamid, Flubendiamide, Triflumuron, Rynaxypyr, Beta-Cyfluthrin, Spirotetramat,
Clothianidin, Thiamethoxam, Thiacloprid, Dinetofuran, Flubendiamide, Cyazypyr, Spinosad, Spinotoram, gamma Cyhalothrin, 4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on, Thiodicarb, Avermectin, Flonicamid, Pyridalyl, Spiromesifen, Sulfoxaflor, Profenophos, Thriazophos, Endosulfan; Cotton Fungicides: Etridiazole, Metalaxyl, Quintozene; Soybean Herbicides: Alachlor, Bentazone, Trifluralin, Chlorimuron-Ethyl, Cloransulam-Methyl, Fenoxaprop, Fomesafen, Fluazifop, Glyphosate, Imazamox, Imazaquin, Imazethapyr, (S-)Metolachlor, Metribuzin, Pendimethalin, Tepraloxydim, Glufosinate; Soybean Insecticides: Lambda-cyhalothrin, Methomyl, Parathion, Thiocarb, Imidacloprid, Clothianidin, Thiamethoxam, Thiacloprid, Acetamiprid, Dinetofuran, Flubendiamide, Rynaxypyr, Cyazypyr, Spinosad, Spinotoram, Emamectin-Benzoate, Fipronil, Ethiprole, Deltamethrin, B-Cyfluthrin, gamma and lambda Cyhalothrin, 4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on, Spirotetramat, Spinodiclofen, Triflumuron, Flonicamid, Thiodicarb, beta-Cyfluthrin; Soybean Fungicides: Azoxystrobin, Cyproconazole, Epoxiconazole, Flutriafol, Pyraclostrobin, Tebuconazole, Trifloxystrobin, Prothioconazole, Tetraconazole; Sugarbeet Herbicides: Chloridazon, Desmedipham, Ethofumesate, Phenmedipham, Triallate, Clopyralid, Fluazifop, Lenacil, Metamitron, Quinmerac, Cycloxydim, Triflusulfuron, Tepraloxydim, Quizalofop; Sugarbeet Insecticides: Imidacloprid, Clothianidin, Thiamethoxam, Thiacloprid, Acetamiprid, Dinetofuran, Deltamethrin, B-Cyfluthrin, gamma/lambda Cyhalothrin, 4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on, Tefluthrin, Rynaxypyr, Cyaxypyr, Fipronil, Carbofuran; Canola Herbicides: Clopyralid, Diclofop, Fluazifop, Glufosinate, Glyphosate, Metazachlor, Trifluralin Ethametsulfuron, Quinmerac, Quizalofop, Clethodim, Tepraloxydim; Canola Fungicides: Azoxystrobin, Carbendazim, Fludioxonil, Iprodione, Prochloraz, Vinclozolin; Canola Insecticides: Carbofuran, Organophosphates, Pyrethroids, Thiacloprid, Deltamethrin, Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dinetofuran, B-Cyfluthrin, gamma and lambda Cyhalothrin, tau-Fluvaleriate, Ethiprole, Spinosad, Spinotoram, Flubendiamide, Rynaxypyr, Cyazypyr, 4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on.
[0091] "Pest" includes but is not limited to, insects, fungi, bacteria, nematodes, mites, ticks, and the like. Insect pests include insects selected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, etc., particularly Coleoptera, Lepidoptera, and Diptera.
[0092] The order Coleoptera includes the suborders Adephaga and Polyphaga. Suborder Adephaga includes the superfamilies Caraboidea and Gyrinoidea, while suborder Polyphaga includes the superfamilies Hydrophiloidea, Staphylinoidea, Cantharoidea, Cleroidea, Elateroidea, Dascilloidea, Dryopoidea, Byrrhoidea, Cucujoidea, Meloidea, Mordelloidea, Tenebrionoidea, Bostrichoidea, Scarabaeoidea, Cerambycoidea, Chrysomeloidea, and Curculionoidea. Superfamily Caraboidea includes the families Cicindelidae, Carabidae, and Dytiscidae. Superfamily Gyrinoidea includes the family Gyrinidae. Superfamily Hydrophiloidea includes the family Hydrophilidae. Superfamily Staphylinoidea includes the families Silphidae and Staphylinidae. Superfamily Cantharoidea includes the families Cantharidae and Lampyridae. Superfamily Cleroidea includes the families Cleridae and Dermestidae. Superfamily Elateroidea includes the families Elateridae and Buprestidae. Superfamily Cucujoidea includes the family Coccinellidae. Superfamily Meloidea includes the family Meloidae. Superfamily Tenebrionoidea includes the family Tenebrionidae. Superfamily Scarabaeoidea includes the families Passalidae and Scarabaeidae. Superfamily Cerambycoidea includes the family Cerambycidae. Superfamily Chrysomeloidea includes the family Chrysomelidae. Superfamily Curculionoidea includes the families Curculionidae and Scolytidae.
[0093] The order Diptera includes the Suborders Nematocera, Brachycera, and Cyclorrhapha. Suborder Nematocera includes the families Tipulidae, Psychodidae, Culicidae, Ceratopogonidae, Chironomidae, Simuliidae, Bibionidae, and Cecidomyiidae. Suborder Brachycera includes the families Stratiomyidae, Tabanidae, Therevidae, Asilidae, Mydidae, Bombyliidae, and Dolichopodidae. Suborder Cyclorrhapha includes the Divisions Aschiza and Aschiza. Division Aschiza includes the families Phoridae, Syrphidae, and Conopidae. Division Aschiza includes the Sections Acalyptratae and Calyptratae. Section Acalyptratae includes the families Otitidae, Tephritidae, Agromyzidae, and Drosophilidae. Section Calyptratae includes the families Hippoboscidae, Oestridae, Tachinidae, Anthomyiidae, Muscidae, Calliphoridae, and Sarcophagidae.
[0094] The order Lepidoptera includes the families Papilionidae, Pieridae, Lycaenidae, Nymphalidae, Danaidae, Satyridae, Hesperiidae, Sphingidae, Saturniidae, Geometridae, Arctiidae, Noctuidae, Lymantriidae, Sesiidae, and Tineidae.
[0095] Nematodes include parasitic nematodes such as root-knot, cyst, and lesion nematodes, including Heterodera spp., Meloidogyne spp., and Globodera spp.; particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode); Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes). Lesion nematodes include Pratylenchus spp., e.g. Pratylenchus penetrans.
[0096] Insect pests of the invention for the major crops include: Maize: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Helicoverpa zea, corn earworm; Spodoptera frugiperda, fall armyworm; Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; Diabrotica virgifera, western corn rootworm; Diabrotica longicornis barberi, northern corn rootworm; Diabrotica undecimpunctata howardi, southern corn rootworm; Melanotus spp., wireworms; Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blot leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant; Tetranychus urticae, twospotted spider mite; Sorghum: Chilo partellus, sorghum borer; Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm; Elasmopalpus lignosellus, lesser cornstalk borer; Feltia subterranea, granulate cutworm; Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid; Blissus leucopterus leucopterus, chinch bug; Contarinia sorghicola, sorghum midge; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, twospotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; Diabrotica undecimpunctata howardi, southern corn rootworm; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly; Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle; Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, cotton budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm; Pectinophora gossypiella, pink bollworm; Anthonomus grandis, boll weevil; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper; Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, twospotted spider mite; Rice: Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhopper; Blissus leucopterus leucopterus, chinch bug; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm; Heliothis virescens, cotton budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips; Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, twospotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Schizaphis graminum, greenbug; Blissus leucopterus leucopterus, chinch bug; Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug; Delia platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite; Oil Seed Rape: Brevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, Flea beetle; Mamestra configurata, Bertha armyworm; Plutella xylostella, Diamond-back moth; Delia ssp., Root maggots.
Methods for Increasing Plant Yield
[0097] Methods for increasing plant yield are provided. The methods comprise providing a plant or plant cell expressing a polynucleotide encoding the pesticidal polypeptide sequence disclosed herein and growing the plant or a seed thereof in a field infested with a pest against which said polypeptide has pesticidal activity. In some embodiments, the polypeptide has pesticidal activity against a lepidopteran, coleopteran, dipteran, hemipteran, or nematode pest, and said field is infested with a lepidopteran, hemipteran, coleopteran, dipteran, or nematode pest.
[0098] As defined herein, the "yield" of the plant refers to the quality and/or quantity of biomass produced by the plant. By "biomass" is intended any measured plant product. An increase in biomass production is any improvement in the yield of the measured plant product. Increasing plant yield has several commercial applications. For example, increasing plant leaf biomass may increase the yield of leafy vegetables for human or animal consumption. Additionally, increasing leaf biomass can be used to increase production of plant-derived pharmaceutical or industrial products. An increase in yield can comprise any statistically significant increase including, but not limited to, at least a 1% increase, at least a 3% increase, at least a 5% increase, at least a 10% increase, at least a 20% increase, at least a 30%, at least a 50%, at least a 70%, at least a 100% or a greater increase in yield compared to a plant not expressing the pesticidal sequence.
[0099] In specific methods, plant yield is increased as a result of improved pest resistance of a plant expressing a pesticidal protein disclosed herein. Expression of the pesticidal protein results in a reduced ability of a pest to infest or feed on the plant, thus improving plant yield.
[0100] The following examples are offered by way of illustration and not by way of limitation.
EXPERIMENTAL
Example 1
Discovery of Novel Pesticidal Genes from Bacillus thuringiensis Having Homology to Pesticidal Genes
[0101] Novel pesticidal genes were identified from the bacterial strains listed in Table 1 using the following steps:
[0102] Preparation of extrachromosomal DNA from the strain, which includes plasmids that typically harbor delta-endotoxin genes
[0103] Mechanical shearing of extrachromosomal DNA to generate size-distributed fragments
[0104] Cloning of ˜2 Kb to ˜10 Kb fragments of extrachromosomal DNA
[0105] Outgrowth of ˜1500 clones of the extrachromosomal DNA
[0106] Partial sequencing of the 1500 clones using primers specific to the cloning vector (end reads)
[0107] Identification of putative toxin genes via homology analysis via the MiDAS approach (as described in U.S. Patent Publication No. 20040014091, which is herein incorporated by reference in its entirety)
[0108] Sequence finishing (walking) of clones containing fragments of the putative toxin genes of interest
TABLE-US-00001
[0108] TABLE 1 Amino Molecular Nucleotide acid Gene weight SEQ ID SEQ ID name Strain (Da) Closest homolog NO NO axmi190 ATX12995 73761 37.9% Axmi143 1 28 axmi191 ATX12995 58498 27.7% Cry36Aa1 2 29 axmi192 ATX12995 90190 64.9% Cry20Aa1 3 30 axmi193 ATX12995 35179 43.7% Mtx2 4 31 axmi194 ATX24031 34428.86 22% Cry55Aa 5 32 axmi195 ATX24031 27107 45.6% Axmi194 6 33 axmi1965 ATX24031 161013 46.2% Cry5Aa1 7 34 axmi196 ATX24031 35% Cry13Aa1_trunc 35 (truncated) axmi197 ATX28233 40196.37 38.2% Axmi019 8 36 axmi198 ATX28233 43908 74.9% Axmi072 9 37 axmi199 ATX28233 40145 60.3% Axmi197 10 38 axmi2001 ATX15076 77098 40.8% Axmi134_trunc 11 39 axmi2012 ATX15076 64361 85.9% Axmi084 12 40 axmi202 ATX12978 72115 17.3% Mtx2 13 41 axmi203 ATX27776 110587 21.5% Axmi148 14 42 axmi203 43 (truncated) axmi204 ATX13053 65491 34.5% Axmi191 15 44 axmi206 ATX27753 63317 28.7% Axmi182 16 45 axmi207 ATX4846 146032 86.8% Axmi134 17 46 axmi207 ATX4846 83.6% Axmi134_trunc 47 (truncated) axmi208 ATX4846 142602 92.1% Axmi134 18 48 axmi208 92.8% Axmi134_trunc 49 (truncated) axmi209 ATX4846 34633 25.3% Axmi180 19 50 axmi210 ATX13028 131083 85.4% Axmi043 20 51 axmi210 80.5% Axmi043_trunc 52 (truncated) axmi2116 ATX13048 137085 74.1% Cry7Ca 21 53 axmi211 59.1% Cry7Ca_trunc 54 (truncated) axmi2127 ATX13003 86972 33.2% Axmi035 22 55 axmi2133 ATX13003 31570 27% Cry15Aa(Bti) 23 56 axmi2144 ATX13003 32786 48.3% Axmi213 24 57 axmi215 ATX13020 148168 60.3% Axmi155 25 58 axmi215 39.7% Axmi155_trunc 59 (truncated) axmi216 ATX13020 41897 28.5% Axmi194 26 60 axmi217 ATX13020 43043 24% Axmi185 27 61 1pairs with axmi201 2pairs with axmi200 3pairs with axmi214 4pairs with axmi213 5Upon examination of the sequence of ATX24031, two overlapping open reading frames (ORFs) were identified, each with homology to endotoxin-like genes. After inspection of these ORFs and their encoded proteins, it was apparent that these two ORFs likely originated from a single ORF that had suffered a single nucleotide insertion (or larger insertion creating a single nucleotide frame-shift) in the region from nucleotide 224 to 309 from the start of the first ORF. These orfs are designated herein as ATX24031_contig4_orf1 (SEQ ID NO: 63) and ATX424031_contig4_orf2 (SEQ ID NO: 64). The full-length sequence is set forth in SEQ ID NO: 65. A composite ORF that has homology to endotoxins over its entirely can be assembled by "fixing" the insertion to create a single ORF. While it is understood that multiple solutions can be created to yield such an ORF and these solutions will differ in the region of overlap between the ORFs, one solution is provided herein, which is designated as axmi196 (SEQ ID NO: 7). 6A p19/CryBP1-like gene was identified immediately upstream of axmi211. The nucleotide sequence for this gene is set forth in SEQ ID NO: 66, and the amino acid sequence is set forth in SEQ ID NO: 67. 6An p19-like gene was identified immediately upstream of axmi212. The nucleotide sequence for this gene is set forth in SEQ ID NO: 68, and the amino acid sequence is set forth in SEQ ID NO: 69.
Example 2
Expression in Bacillus
[0109] The pesticidal gene disclosed herein is amplified by PCR from pAX980, and the PCR product is cloned into the Bacillus expression vector pAX916, or another suitable vector, by methods well known in the art. The resulting Bacillus strain, containing the vector with axmi gene is cultured on a conventional growth media, such as CYS media (10 g/l Bacto-casitone; 3 g/l yeast extract; 6 g/l KH2PO4; 14 g/l K2HPO4; 0.5 mM MgSO4; 0.05 mM MnCl2; 0.05 mM FeSO4), until sporulation is evident by microscopic examination. Samples are prepared and tested for activity in bioassays.
Example 3
Insecticidal activity of Axmi-191 and Axmi-192
Gene Expression and Purification
[0110] The DNA regions encoding the toxin domains of axmi-191 and axmi-192 were separately cloned into an E. coli expression vector pMAL-C4x behind the malE gene coding for Maltose binding protein (MBP). These in-frame fusions resulted in MBP-Axmi fusion proteins expression in E. coli.
[0111] For expression in E. coli, BL21*DE3 was transformed with individual plasmids. Single colony was inoculated in LB supplemented with carbenicillin and glucose, and grown overnight at 37° C. The following day, fresh medium was inoculated with 1% of overnight culture and grown at 37° C. to logarithmic phase. Subsequently, cultures were induced with 0.3 mM IPTG for overnight at 20° C. Each cell pellet was suspended in 20 mM Tris-Cl buffer, pH 7.4+200 mM NaCl+1 mM DTT+ protease inhibitors and sonicated. Analysis by SDS-PAGE confirmed expression of fusion proteins.
[0112] Total cell free extracts were run over amylose column attached to FPLC for affinity purification of MBP-axmi fusion proteins. Bound fusion protein was eluted from the resin with 10 mM maltose solution. Purified fusion proteins were then cleaved with Factor Xa to remove the amino terminal MBP tag from the Axmi protein. Cleavage and solubility of the proteins was determined by SDS-PAGE.
Insect Bioassays
[0113] Cleaved proteins were tested in insect assays with appropriate controls. A 5-day read of the plates showed that Axmi191 and Axmi192 had activity against diamondback moth species. Axmi 191 showed stunting and Axmi192 showed severe stunting and 100% mortality.
Example 4
Construction of Synthetic Sequences
[0114] In one aspect of the invention, synthetic toxin sequences were generated. These synthetic sequences have an altered DNA sequence relative to the parent toxin sequence, and encode a protein that is collinear with the parent toxin protein to which it corresponds, but lacks the C-terminal "crystal domain" present in many delta-endotoxin proteins.
[0115] In another aspect of the invention, modified versions of synthetic genes are designed such that the resulting peptide is targeted to a plant organelle, such as the endoplasmic reticulum or the apoplast. Peptide sequences known to result in targeting of fusion proteins to plant organelles are known in the art. For example, the N-terminal region of the acid phosphatase gene from the White Lupin Lupinus albus (Genebank ID GI:14276838; Miller et al. (2001) Plant Physiology 127: 594-606) is known in the art to result in endoplasmic reticulum targeting of heterologous proteins. If the resulting fusion protein also contains an endoplasmic retention sequence comprising the peptide N-terminus-lysine-aspartic acid-glutamic acid-leucine (i.e. the "KDEL" motif (SEQ ID NO: 70) at the C-terminus, the fusion protein will be targeted to the endoplasmic reticulum. If the fusion protein lacks an endoplasmic reticulum targeting sequence at the C-terminus, the protein will be targeted to the endoplasmic reticulum, but will ultimately be sequestered in the apoplast.
Example 5
Assays for Pesticidal Activity
[0116] The ability of a pesticidal protein to act as a pesticide upon a pest is often assessed in a number of ways. One way well known in the art is to perform a feeding assay. In such a feeding assay, one exposes the pest to a sample containing either compounds to be tested, or control samples. Often this is performed by placing the material to be tested, or a suitable dilution of such material, onto a material that the pest will ingest, such as an artificial diet. The material to be tested may be composed of a liquid, solid, or slurry. The material to be tested may be placed upon the surface and then allowed to dry. Alternatively, the material to be tested may be mixed with a molten artificial diet, then dispensed into the assay chamber. The assay chamber may be, for example, a cup, a dish, or a well of a microtiter plate.
[0117] Assays for sucking pests (for example aphids) may involve separating the test material from the insect by a partition, ideally a portion that can be pierced by the sucking mouth parts of the sucking insect, to allow ingestion of the test material. Often the test material is mixed with a feeding stimulant, such as sucrose, to promote ingestion of the test compound.
[0118] Other types of assays can include microinjection of the test material into the mouth, or gut of the pest, as well as development of transgenic plants, followed by test of the ability of the pest to feed upon the transgenic plant. Plant testing may involve isolation of the plant parts normally consumed, for example, small cages attached to a leaf, or isolation of entire plants in cages containing insects.
[0119] Other methods and approaches to assay pests are known in the art, and can be found, for example in Robertson, J. L. & H. K. Preisler. 1992. Pesticide bioassays with arthropods. CRC, Boca Raton, Fla. Alternatively, assays are commonly described in the journals "Arthropod Management Tests" and "Journal of Economic Entomology" or by discussion with members of the Entomological Society of America (ESA).
Example 6
Expression of pMal Fusion Proteins
[0120] For expression in E. coli, select genes of the invention were cloned into a pMal expression vector (New England Biolabs) such that the protein was expressed in E. coli with an N-terminal fusion to maltose binding protein (MBP). A nucleotide sequence encoding a truncated variant of Axmi207 (corresponding to positions 15 through 647 of SEQ ID NO:47) was used to test for bioactivity. The truncated variant Axmi207 sequence is set forth in SEQ ID NO:62. For Axmi196, Axmi204, and Axmi209, the full length native sequence was used.
[0121] These fusion proteins were then purified by affinity chromatography as known in the art. The purified proteins were then cleaved with protease as known in the art to separate the MBP from the protein of the invention. The resulting proteins were then tested in bioassays against selected pests. The results are shown in Table 2.
TABLE-US-00002 TABLE 2 axmi207 (truncated Pest axmi196 axmi204 variant) axmi209 DBM Severe stunt Severe stunt, Severe stunt, 100% 100% 100% mortality mortality mortality CPB 100% mortality SWCB Moderate Severe stunt, Strong stunt, stunt, 50% 100% 50% mortality mortality mortality VBC Stunted Stunted ECB Moderate stunt, 50% mortality Hz Stunted Stunted FAW Stunted SCB Strong stunt Stunted SCN 100% mortality C. elegans 100% mortality Pratylenchus 40% Penetrans mortality DBM--Diamondback moth CPB--Colorado potato beetle SWCB--Southwestern corn borer VBC--Velvetbean caterpillar ECB--European corn borer Hz--Helicoverpa zea FAW--Fall armyworm SCB--Sugarcane borer SCN--Soybean cyst nematode
Example 7
Vectoring of the Toxin Genes of the Invention for Plant Expression
[0122] Each of the coding regions of the genes of the invention is connected independently with appropriate promoter and terminator sequences for expression in plants. Such sequences are well known in the art and may include the rice actin promoter or maize ubiquitin promoter for expression in monocots, the Arabidopsis UBQ3 promoter or CaMV 35S promoter for expression in dicots, and the nos or PinII terminators. Techniques for producing and confirming promoter-gene-terminator constructs also are well known in the art.
Example 8
Transformation of the Genes of the Invention into Plant Cells by Agrobacterium-Mediated Transformation
[0123] Ears are collected 8-12 days after pollination. Embryos are isolated from the ears, and those embryos 0.8-1.5 mm in size are used for transformation. Embryos are plated scutellum side-up on a suitable incubation media, and incubated overnight at 25° C. in the dark. However, it is not necessary per se to incubate the embryos overnight. Embryos are contacted with an Agrobacterium strain containing the appropriate vectors for Ti plasmid mediated transfer for 5-10 min, and then plated onto co-cultivation media for 3 days (25° C. in the dark). After co-cultivation, explants are transferred to recovery period media for five days (at 25° C. in the dark). Explants are incubated in selection media for up to eight weeks, depending on the nature and characteristics of the particular selection utilized. After the selection period, the resulting callus is transferred to embryo maturation media, until the formation of mature somatic embryos is observed. The resulting mature somatic embryos are then placed under low light, and the process of regeneration is initiated as known in the art. The resulting shoots are allowed to root on rooting media, and the resulting plants are transferred to nursery pots and propagated as transgenic plants.
Example 9
Transformation of Maize Cells with the Toxin Genes of the Invention
[0124] Maize ears are collected 8-12 days after pollination. Embryos are isolated from the ears, and those embryos 0.8-1.5 mm in size are used for transformation. Embryos are plated scutellum side-up on a suitable incubation media, such as DN62A5S media (3.98 g/L N6 Salts; 1 mL/L (of 1000× Stock) N6 Vitamins; 800 mg/L L-Asparagine; 100 mg/L Myo-inositol; 1.4 g/L L-Proline; 100 mg/L Casaminoacids; 50 g/L sucrose; 1 mL/L (of 1 mg/mL Stock) 2,4-D), and incubated overnight at 25° C. in the dark.
[0125] The resulting explants are transferred to mesh squares (30-40 per plate), transferred onto osmotic media for 30-45 minutes, then transferred to a beaming plate (see, for example, PCT Publication No. WO/0138514 and U.S. Pat. No. 5,240,842).
[0126] DNA constructs designed to express the genes of the invention in plant cells are accelerated into plant tissue using an aerosol beam accelerator, using conditions essentially as described in PCT Publication No. WO/0138514 After beaming, embryos are incubated for 30 min on osmotic media, then placed onto incubation media overnight at 25° C. in the dark. To avoid unduly damaging beamed explants, they are incubated for at least 24 hours prior to transfer to recovery media. Embryos are then spread onto recovery period media, for 5 days, 25° C. in the dark, then transferred to a selection media. Explants are incubated in selection media for up to eight weeks, depending on the nature and characteristics of the particular selection utilized. After the selection period, the resulting callus is transferred to embryo maturation media, until the formation of mature somatic embryos is observed. The resulting mature somatic embryos are then placed under low light, and the process of regeneration is initiated by methods known in the art. The resulting shoots are allowed to root on rooting media, and the resulting plants are transferred to nursery pots and propagated as transgenic plants.
Materials
TABLE-US-00003
[0127] DN62A5S Media Components per liter Source Chu'S N6 Basal 3.98 g/L Phytotechnology Labs Salt Mixture (Prod. No. C 416) Chu's N6 Vitamin 1 mL/L (of 1000x Stock) Phytotechnology Labs Solution (Prod. No. C 149) L-Asparagine 800 mg/L Phytotechnology Labs Myo-inositol 100 mg/L Sigma L-Proline 1.4 g/L Phytotechnology Labs Casaminoacids 100 mg/L Fisher Scientific Sucrose 50 g/L Phytotechnology Labs 2,4-D (Prod. No. 1 mL/L (of 1 mg/mL Stock) Sigma D-7299)
[0128] Adjust the pH of the solution to pH to 5.8 with 1N KOH/1N KCl, add Gelrite (Sigma) to 3 g/L, and autoclave. After cooling to 50° C., add 2 ml/L of a 5 mg/ml stock solution of Silver Nitrate (Phytotechnology Labs). Recipe yields about 20 plates.
[0129] All publications and patent applications mentioned in the specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
[0130] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.
Sequence CWU
1
1
7011922DNABacillus thuringiensis 1ttggaaatta aaattggaag cggaggaaca
tatatgaatc catataacaa cgaaagctat 60gaaattattg atttaaatac ttcaccttat
ccttctaaca gaaataattc taggtatcct 120tatgcaaatg cttgcggttt tccagaaaac
gtagattgga cagcaggcgc aagtgcaatg 180ataattgtag ctggtacttt attaagtgct
ataggttcag gaggagtagg tatagttgct 240gcaggtatta tatcagttgg tacactattc
cctttttttt ggcctcaaga taagcctact 300gcacaagtat ggaaagattt tattaaacaa
ggggatacta taactaataa aacaatatca 360gcagccgtag aatctctagt actcgcagaa
ttaaatggtt taaaatctat acttgatgtt 420tatactgatg ctttagaact ttggaaaaaa
gataaaaata atatagtcaa tagagacaat 480gtaaaaagta tttttacaaa tttacatcta
caatttgtag ccgctatgcc aaaatttgca 540acaaatggtt atgaagtaat attattatct
acttacacag cagctgcact tcttcatatt 600acttttttac atgaagctct tcaatatgca
aatgaatgga atttagctcg aagtgaagga 660accttctatc gtggacaatt aattcaagca
atagaaaact acattaatta ttgtgaaaaa 720tggtatcgtg aaggtttaga gatacttaaa
aattctactt gggatatata tgctgcgtat 780caaaatgaat acactctaag tatattaaat
gttatttcaa tttttccaag atttgatata 840cgtaatttcc ctacaaatat agcaactcga
ttagaatcta cacaaaaact ttatacaaca 900acaccaaata tgaaagcatt aaaaacaaat
aactcaattg attatataaa agataaactt 960atacctcctt tagatttatt taaaaaatta
aaaagtttaa ctttttatac atttttagat 1020agcaataacc aatatgatca tttacaaggt
attgtaaata atagttatta tactaatatt 1080tccactaaca aaatcttttc ttctggaact
accgaaggta gttcatatca actaggtttg 1140gcttctgatc aagttattta ctacactgac
atcttccatc atctaaatca aagtaatttt 1200aaggatggtt cccttggaat taaaataatt
aattttaata ttataaataa atataatgag 1260gtttctcaaa aatcttatga ttctaatgca
acaagtaatc taatactaga agttatatta 1320ccttttctaa aaacaactga gaaagattat
aaatatattt tatcttatat tacaataact 1380ccacagcaga tagtaggatg tctaagtcct
agttatatat atggatttat ttggacacat 1440agtagtgtta atcttaacaa tactattcat
tatacaaata aaaataattt ttctcaaatt 1500acacaaattt ctgcagtaaa agcatatctg
aaaaaagatc gagtttcagt tatagaagga 1560ccaggtcata caggcggaga tttagttaaa
tttacacaat gggatgattc aatttcaact 1620cattatcaat ttactagcag tggtgaatat
aaaatacgtg tccgatatgc ttctactgct 1680caagttaatc aaaccagcgg acttagtatg
acgatatacc ataaaggaaa tcctacagaa 1740acatgggatt taaacataaa taacaaatca
gatacaatac ttaatttaaa tgaaccaaaa 1800tacaatcatt ttcaatacac agaatttcca
aataaaactc ttataataaa taaagaccca 1860aattctccat acttagaact aagaatagac
ttaagctata aaggaaatac tgcaacaact 1920ct
192221518DNABacillus thuringiensis
2atgaatggaa atggaaaaca tgataactgg aatcacaatc aacaaatatc aaatgtccaa
60atgaaccaca atcatggtag atcatatgat tgcagttgtc aacaaaatca gtatgggtat
120gagcaacaaa aacagcagta tgaacagaat aacagccaat atatgcagaa taatctggga
180aacgaaaata ggaatggatt gtatccttat caagaaaatc aatatgaaca aaataagaat
240tattatgcat caaacaattt aacatataat cagtctgatt tgtataattc taatcctcaa
300aatatgtata aacatcaaac atattctaat gatttttatt gttctcctag ctatacagca
360ggtgaaaata atatattaga tctattaggt acagaaagta aacaattcca aaaaatttca
420aatataaata ctaaagattt acatcgaagt ataactgcga gcaatactca aattggttat
480caaattgata ctcgtgttcc aggaccatgt aaaggtgtag attatcaaaa cacagtaacc
540tatgaacaaa attcaatagg tggcgattcc caatacttga ttttttataa aacggattat
600actgatgcat ttattattgc gaatagagca aatggtcgag ttttagaagt aatacctagt
660tcagttaatg gttttgtaac aatttctaat atgtttactt ataatcaaaa tcaacttttt
720attcgtacta aaatatcaaa taatgataat tcagatgatg ttccattttc tttaacaaca
780gaaaacaatc aaacattaaa catatgccat catgaatttc aatataatac taaaattaca
840gctcttgata atgcatatcg tttggatgat aaggttttat ttaaaccaac tagagataaa
900atcaacatat catttccaaa tatggtagtg aatgcgaagg agaaattacc agaacccgag
960gaattaacaa atatggataa gaatactctt tttataccga aagtgattat aagtaaaacg
1020ttaattccag gtataattgt aaacgatgta actttattaa aggagcaaca aatagcaaaa
1080agtccatatt atgtattaga atatgttcaa tcttgggaag aagtgtataa tgaaatagta
1140cctgcttata gaccttcgta tacttggact tcaacagatg gaattagaca cgttaatcta
1200ttagatataa agaatactat aaatatatca ataggtggaa ctagtcaagg ctggggatta
1260agatttagtg ataaatcaga tctttttaaa aacataatta catcagcatt cattataaaa
1320tcaacacaag ctccagatat gggattcagt gagaatgata tagatcagta ctatggtaag
1380aatattgaca gtagagttaa aatatatata aaaacccata atttaatatt aagacgttta
1440gatcaattga acaattcaat agctacatgg acaatatttg agaatacaaa acctgttata
1500agaacgtttc caattagt
151832337DNABacillus thuringiensis 3atgaaggata aaaaatactg gaaatacgag
ggaggaacca aaatgaatcc ttatcagaat 60aagaatgaat atgaaatagt aaataatccg
caaaattata atactgtttc caatagatat 120ccttacacga atgatccaaa tgttgcaata
caaaatacga attataaaga ttggatgaat 180gggtatgaag aaattaatcc ttcttcaata
tcgttaattt tagcttcaat aggaattctt 240aatcaagcaa ttgctttaac tggagtatta
ggtaagacac cagaaattat taacatagta 300caagaaatgg tgggattaat tagcgggagt
acaggcaatg atttattagt acatacagaa 360caacttattc aacaaacttt agcacaacag
tatagaaacg cagcaaccgg agcggtgaat 420gctatatcta aatcatacaa tgattatttg
atgtttttta ggcaatggga acgtaataga 480acttctcaaa atggactaca agtagagagt
gcttttaata ctgttaatac tttatgtctt 540cgtactttaa ctcctcagga agcactttct
cgcagaggat ttgaaactct tttattacca 600aactatgcac tagcggcaaa tttccatttg
ttattattaa gagatgctgt tctttataga 660actcagtggt tacctaattt tatttcaact
acaaatgcga atattgaaat attggaaagg 720tccataaatc aatatcgtaa tcattgtaat
cattggtaca atgacggctt aaatagattt 780gcacgtacat cttttgatga ttgggttcgg
tttaatgctt atcgtagaga tatgacgtta 840tcggtattag attttgttac agtatttcca
acttataatc ctataaactt tccaacacca 900acaaatgttg aattgactag aatcgtttat
accgatccaa taagtccacc tagaggatat 960gcaagaactg gctcacctag ttttcgtcaa
atggaagatc taattatttc cggtagccct 1020agtttcttga atcaattaag tatatttaca
acttattatc atgatcctcg taatgtaaat 1080agagactttt gggccgggaa tcggaattat
ttaagcaatg ggacttctcg acagtctgga 1140gctaccacac cttggcgaac taatatacct
atgcaaaaca ttgatatttt cagagtaaat 1200ctaactactc atgacattga tgatatatca
cgaagttatg gaggagttca tagatctgat 1260ttcattggtg taaatacaat aaataatcaa
agaacaacat tgttctatca ccaaaatgtg 1320gatacttccc gttttctaat aaggaatgaa
acagtatttt taccagggga ttccggctta 1380gcaccaaatg aacgtaatta tactcacagg
ttatttcaag tgatgaccac atatcgtact 1440aacccgaatg ctcgtagggc agctttttta
catgcatgga cgcatagaag tttaagacgt 1500agaaatggat ttaggacgga tcagattatg
caaatacctg ctgtgaagag cataagtaat 1560ggtggtgatc gtgcagtcat atcctatact
ggagaaaata tgatgaaatt agataactta 1620actgcaagtt tatcctataa attaacagcg
gaggattccg aagcatcgaa tacacgtttt 1680atagtgcgta ttcgttatgc tagtatgaac
aataatagat tgaatcttat tttaaatggt 1740actcagatag catcgctgaa tgtggaaggt
acaatgcaaa atggcggatc attaacaaat 1800cttcaatctg aaaattttaa atatgctaca
ttttcaggta atttcaagat gggttctcag 1860tctatagtag gtatttttaa agagatatct
aatgcagact ttattttaga taaaattgaa 1920ttgattccaa ttcattttat gccattatta
gaacaaaaac aaagctacaa caattacgac 1980caaaacatgg atactacata tcaaccaaac
tatgacactt ataatcaaaa tgccaatggt 2040atgtatgacg atacatacta tccaaataat
aatgatagtt ataatcaaaa taataccgat 2100atgtatgatt caggctacaa taacaaccaa
aatactaact ataattatga tcaagaatat 2160aatacttaca atcaaaatat ggaaaatacg
tatgaccaat cgtatgaaaa ttacaatcca 2220gaaaccaaca attacaacca ataccctaat
gatatgtaca atcaagagta tactaacgac 2280tacaaccaaa actccggctg caggtgtaac
caagggtata ataataatta ccctaaa 23374966DNABacillus thuringiensis
4atgaatttct tgtacaattt tgtgacatta gatatgctaa tattaaatag attagaagga
60agtgatttta aaatgaagaa aaaagcaata gtgtgtggct tactagctag tactttatta
120ggcggcggta cttttgtaga tgctgtgagt gcggctgaaa ttcaaaaaac taatcattta
180aacaagtatg atagtgcaca agagaaagct ctacaagata ttaaccaaga agcgttacaa
240gatattgatc aaaaagtcaa taagatgatt gattctatcc cacccatttt tggatcaaaa
300tatacacgta cagatcgcta cggtgaaagt cttacttatt caggaataaa tctaaaagaa
360aataatagta caaatgttga accgatgtac tttggttcaa atacatttta taacgataca
420gagctagaac aatcctataa cactacttct tttagtgaag ctgttactaa atcaactact
480actcaaacac aaaatggatt taaatcaggt gtaactacag gaggaaaggt tgggatacct
540tttgtagctg aaggtgaagt gaaaatcaat cttgaatata attttacaca caccaattca
600aatactacta gcaagactac aactttaaca gcccctccac aacctgttaa ggttcctgca
660ggtaaagttt ataaagcaga cgtttatttt gaaaagaaat ctacttcggg tacggttgaa
720ctttatggag atctccttac aggtgtagta gccgagggaa ggacatcatt tgtaggtaac
780gtattacata aggcaactga tacacaaggg ctaattcaat ctcctgagga ttcaaataaa
840gttcgtgcgg ttggaaaagg aacgttcacc actgaacatg gctcaaactt tatcgtcaaa
900acatatgatg taacatcagg gcaaaaatcc gcaaaattgg tagatactag agtaatacct
960ataaaa
9665969DNABacillus thuringiensis 5atgaatttaa ggaggatttc tatgagagtt
tataaaaaat tagcaacgtt ggcacctatt 60gctgcattaa gtacatccat tttatgttct
cctgcaatga catttgcagc agaaaaagaa 120tcaacagtga aacaaaccac acaacaaagt
gcggttcaac aaggtcgtat cattcaaggg 180tatcttatta aaaatggtgt gaaaatacca
gtttatacag gtggattagt aacaaataag 240gctgaacaag gtgcagcagc atttccacaa
ttgtcttcca atcctaatga tcctatccct 300caaaaaggtt ctatttcatc tgaagatgga
aatattggag atattttata tttttctaaa 360actccaatgg gagataatgt ttatataaaa
aaacttgaga ataacaacat tgaaattgga 420aaatataatc gaggcacttt agaattatcg
aaatttgtga cagttaatgg agatccacag 480ggacctataa tgttatttga tgctacggta
aaacgtgaaa cggcatttga aaaaattggt 540ggtgctgtac aaccaaaggc aacacaatat
acttttagtc aagcggtaac atccggttta 600tcaacatcag atgcgattgg cggttcatta
acattaggat ataaaatatc gcttaaagaa 660ggtggcggag tagtaccagc cgaagcaaca
caggaattta gtacacaatt aagtgctaca 720tataatcata caattacagt gacaaaccaa
acaacgaata cacaaacaca aacctttaaa 780cctatagaca gttatggaca atcaacctat
gcagctgctg tgtatcaatt aaaatctcat 840tatacggtga ttccaggagc aggattacag
aagggattaa atagtggata tgtgttagat 900caaacagcgt tttcatatag cgattctgat
ttatatctag ctgtaacacc aggagcaggt 960tcaaatgta
9696756DNABacillus thuringiensis
6ttgatcacta atcaagcagc acaagcaagt gatgcaccct atccagaatt accatcgaat
60ccgaatgatg cgattccaaa tgccggggcc actcatgctg aaaatggaag tgtaggttcg
120gttttatatt ttaaacagat agatttaaat aatctgggag ctggcatagg aaatagtcaa
180aaagattatg tttacgtaga aaaaaaaggt gattctggat atgaattagg aaattacaat
240ccgttaactt tacaaagaac taaaattaaa gattatgata aatccagtga acttgcagaa
300aagatggatg gctattttaa aagtacaatt acacgagata ctttttttag taaaatcgga
360tctggtgtcg taccaaaaaa tgcagcgtat acctttagtc aagcagttac atccggttta
420actacatcag atgcgattgg gggtgcgcta acactcggat ataaagtaag tgttacagaa
480ggcggaggaa tattcccagc tgcagcgtca gaagaattta gtgcacaatt aacagcaact
540tataatcata cgattactgt ttccagccaa gtaacaaata ctcaaacatt gggcattaca
600aaagctgcag atggctatca atatgataaa tatgtaggcg ctgtatatca attgcattcc
660aagtatacat ttaaacctag tgatgaatta caatttgcaa tgaattcacc ttttggatat
720aaggtaattc ttaatcaacg agcacaatca ttccaa
75674323DNABacillus thuringiensis 7atgacaacaa taaatgaatt atatccggct
gtaccttata atgtactggc atatgctcca 60ccacttaatt tagctgattc gacaccatgg
ggtcaaatag ttgttgctga tgcaattaaa 120gaagcttggg ataattttca aaaatatggt
gtattagatt taacagctat aaatcaaggg 180tttgatgatg caaatacagg ttcttttagt
tatcaagctt taatacaaac tgttttgggt 240attataggta caattggtat gacagttcct
gtggctgctc catttgcagc tacagcgcct 300attattagtt tatttgtagg atttttttgg
cctaaaaaag ataagggacc acaattaatc 360gatataattg ataaagaaat taaaaaatta
ttagataagg aattaggaga gcaaaaacgt 420aatgatttag ttagtgcttt aaatgagatg
caagagggag caaatgagtt aagtgatatt 480atgactaatg cactttttga aggtactata
cagggaaatg ttgttactaa tgataaccct 540caaggtaaaa ggcgaactcc taaagctcca
acagttagtg attatgagaa tgtttattcg 600gcatattttg tggaacatgt ggattttaga
aacaaaatat ctacgtttct tactggttct 660tatgatctta tagcactccc attatatgca
ttagcaaaaa caatggagct ttcattgtat 720caatcattta ttaattttgc taataaatgg
atggattttg tatatacaaa agcaattaat 780gaatcagcaa ctgatgatat gaaaagagat
tatcaagcga gatacaatac tcaaaaaagt 840aatttagctg tacaaaaaac acaattgatt
aacaaaatta aagatggtac agatgctgtt 900atgaaagttt ttaaagatac caataattta
ccttcaatag gtactaataa attagcagta 960aatgctcgta ataagtatat tagggcctta
caaataaatt gtttagattt agttgctttg 1020tggcctggct tatatccaga tgaatatctt
ttaccattac aattagataa aacacgtgtt 1080gtattttctg atacaatggg acctgatgaa
acacatgatg gtcaaatgaa agttttaaat 1140atattagact caactacaag ttataaccat
caagatatag gaataagtac aactcaagat 1200gtaaattctt tattatttta tccaagaaaa
gaactgttag aattagattt tgctaaatat 1260atttcatcta gtagtcgttt ttgggtttat
ggatttggct taaaatattc agatgataac 1320ttttatagat atggtgataa cgatccaagc
agtgatttta aacctgcata taagtggttt 1380acgaaaaatt cccagttcga aaaccttcct
acttatggaa atcctactcc tattactaat 1440ttaaatgcta aaactcaagt aacttcttat
cttgatgcat taatatatta tatagacgga 1500ggaactaatc tatataataa tgcgattctt
catgatacag ggggttatat tccgggatat 1560ccaggtgtag aaggatatgg tatgagtaat
aatgaacctt tagcaggaca aaaattaaat 1620gctttatatc ctataaaagt ggaaaatgta
agtggttcac aaggaaaatt aggaacaata 1680gcagcttatg ttcctttaaa tttacaacca
gaaaatatta ttggtgatgc tgatccgaat 1740acaggttttc cccttaatgt aattaaagga
tttccatttg aaaaatatgg acctgattat 1800gagggacgag gaatttcggt tgtaaaagaa
tggataaatg gtgcaaatgc tgtaaaattg 1860tctccaggtc aatcagttgg ggtacaaatt
aaaaatataa caaaacaaaa ttatcaaatt 1920cgtactcgtt atgcaagtaa taacagtaat
caagtatatt ttaatgtaga tccaggtgga 1980tcaccattat ttgcacaatc agtaacattt
gaatctacaa caaatgttac aagtggccaa 2040caaggcgaaa atggtagata tacattaaaa
actatttttt ctggtaatga tctacttaca 2100gtagaaatcc ctgttggaaa tttttatgtg
catgttacga ataaaggatc ttctgatatc 2160tttttagatc gtcttgagtt ttctacagtt
ccttcatatg ttatatattc aggtgattat 2220gatgctacag gtacagatga tgtcttattg
tcagatccac atgagtattt ttatgatgtc 2280atagtgaatg gtactgctag tcattctagt
gcagctactt ctatgaattt gctcaataaa 2340ggaaccgtag taagaagcat tgatattcca
ggtcactcaa cgtcttattc tgtacagtat 2400tcagttccag aaggatttga tgaagttaga
attctcagtt ctcttccgga tattagtgga 2460actataagag tagaatctag taaaccacct
gtatttaaga atgatggtaa tagtggtgat 2520ggtggtaata ctgaatataa ttttaatttt
gatttatcag gattgcaaga tactgggctt 2580tattctggta aacttaaatc tggtattcgt
gtgcaaggta attacactta cacaggtgct 2640ccatctttaa atctggttgt ttacagaaat
aatagtgttg tatccacttt tccagtaggt 2700tctccttttg atatcactat aacaacagaa
actgataagg ttatcctttc attacaacct 2760caacatgggt tggcaacagt tactggtact
ggcacaataa caattcctaa tgataaatta 2820gcaattgttt atgataagtt atttaaatta
ccacatgatt tagaaaatat aagaatacaa 2880gtaaatgcat tattcatatc gagtacacaa
aatgaattag ctaaagaagt aaatgaccat 2940gatattgaag aagttgcatt gaaagtagat
gcattatcgg atgaagtatt tggaaaagag 3000aaaaaagaat tacgtaaact ggtcaatcaa
gcgaaacgtt taagtaaagc acgaaacctt 3060ctggtaggag gcaattttga taattgggaa
gcttggtata aaggaaaaga agttgcaaga 3120gtatctgatc atgaattatt gaagagtgat
catgtattat taccgcctcc aactatgtat 3180ccatcctata tatatcaaaa agtagaagaa
acaaaattaa agccaaatac tcgttatatg 3240atttctggtt tcatcgcaca tgcggaagat
ttagaaattg tggtttctcg ttatgggcaa 3300gaagtaagga aaatagtgca agttccatat
ggagaagctt tcccattaac atccaatgga 3360tcaatttgtt gtacaccaag ttttagacgt
gatggaaaac tatcagatcc acatttcttt 3420agttatagta ttgatgtagg tgaactggat
atgacggcag gtccaggtat tgaattggga 3480cttcgtattg tagatcgatt aggaatggcc
cgtgtaagta atttagaaat tcgtgaagat 3540cgttctttaa cagcaaatga aatacgaaaa
gtgcaacgta tggcaagaaa ttggagaacc 3600gaatatgaga aagaacgtgc agaagtaaca
gcattaattg aacctgtatt aaaccaaatc 3660aatgcgttat atgaaaatgg agattggaat
ggttctattc gttcagatat ttcgtactac 3720gatatagaat ctattgtatt accaacatta
ccaagattac gtcattggtt tgttcctgat 3780atgttaactg aacatggaaa tatcatgaat
cgattcgaag aagcattaaa tcgtgcttat 3840acacagctgg aaggaaatac actattgcat
aacggtcatt ttacaacaga tgcggtaaat 3900tggatgatac aaggagatgc acatcaggta
atattagaag atggtagacg tgtattacga 3960ttaccagact ggtcttcgag tgtatcccaa
acaattgaaa tcgagaaatt tgatccagat 4020aaagaataca acttagtatt tcatgcgcaa
ggagaaggaa cggttacgtt ggagcatgga 4080gaaaaaacaa aatatataga aacgcataca
catcattttg cgaattttac aacatcacaa 4140agtcaaggaa ttacgtttga atcgaataag
gtgaccgtgg aaatttcttc agaagatggg 4200gaattattgg tagatcatat cgcacttgtg
gaagttccta tgtttaacaa gaatcaaatg 4260gtcaatgaaa atagagatgt aaatataaat
agcaatacaa atatgaataa tagcaataat 4320caa
432381092DNABacillus thuringiensis
8atgttaataa aggagatgca atatatgcat tctattaaaa aatataaaaa ggttctatta
60attgcaccac ttgcttgtat gttaacaggt gctattttac ctacagctac tacagttcat
120gcacaagagg tagaaaataa aaaagctgta tcaatgatga agccgggagg agagtttgga
180gcaactaaat attcaaaaga aaatttagta aaggaaatca atcttagatt attaacagcg
240cttgatcgtt caacaagttt gcgtgaaaaa tttcatataa agggcaacga agttttagat
300gttagtcagc ttgatgacac atctaaacaa ttaatggaga aattacaatt aacagctgaa
360ggatcaattg atgtgaaacc acatgtcgat agctataaag atcttggtca aaccaatatt
420gttacatata acaatgataa cggagtggtt ggacagacat ataacacacc agaaacaaca
480gtaaaagaat ctgaaactca tacctactcg aatacagaag gggtgaaatt aggactcgag
540gtaggaacaa aaattacagt tggaattcca tttatcggaa aagatgaaac agaaataaaa
600gcaacatccg aattttctta tgaacataat gattcacaaa caaaaacgaa agaaactgat
660gttacgttta aatcccaacc agtagttgcc gctccaggtg gaacaaccac ttattatggt
720gatattaaaa cagcaacatt ttctggatca tttcaaagtg atgcttatgt agcaggcggt
780ttcgaattga aagttcctat cgcacatgat atggcttcgc caaaaataga tcgttatgaa
840acggctacgc tgacagctgc agatatatat gaaattttta atgcttctaa tgcgatagca
900gcaccaaatt acttaaaact tgataacgca ggtaaaaaag ttcttcttac agacaaagca
960acttttgata taaatggaca aggtggtttt tatacaacat tacaggttaa atttgttcct
1020aaagattcta ataaaaagcc tcaaatgatg tcttataaag aatatgtaca aaaaatgaac
1080aataatgaat ta
109291182DNABacillus thuringiensis 9atgtattcta ttaaaagata taaaaaggta
gcaatagtag ccccacttgt ttgtttattg 60ggaacgggac taacatttgt taataaacca
ataccagctg ctgcggcagt aactacaaat 120tattctacag cagattctgc atcaaatttc
caacccatta gtaaatatac tttagccgga 180gatctatatg aacgatatat gagagctcta
gtaagacatc ctgaattact ttcttcaggt 240ggtttaaaac cagtaactaa tcaaacggat
ctagaacaaa tcgacggata ctacaaggta 300atggctcaat tcataagaga caataatcag
aattttccat ctccttttaa tagaccaagt 360atgaaattga tgactggagt taatccgttt
tttaattggg ctcctcaata tactaatctt 420tctactcaaa atgtaattaa cttagataat
ccaaaagtag atgattataa agaagataat 480attgaactag ctacttatac taataacaca
acatcagaac aaactttttc gacgccttca 540aaatcagaaa aagtaacaga ttctttcaca
tattctaatt cagaaggtgg aaaattagga 600gtttcttcca cgactacaat tagagcggga
attccaatag cgcaagctca agaaactctt 660acaatgtcat ttgaagcaac ttataatcat
acaagctcga atacatcttc tactgaaaag 720acagttacat atccatctca agtactaaag
tgcctaccag gatatagaac ttctttaatt 780gtaaaagtat ctcaagcgaa tttttctggt
acaatggatt ttgacgttga accaactgtg 840agttcattaa tagatggtat agaaaaaaat
tggaaagaca taaaagacga taagacaata 900aaaggagata aaagtggaga ttacacagtc
ccaaatcgac aagaattttt atataatgtg 960tataaatatt cagatttacc aattccatct
tatgttaaat tagatgataa aaagaaaact 1020gtatcatttg gaaaagttac aactccatat
acaggtgtag caggtcattt atcagaagca 1080aatgcaacac aagtaaaact ggaatcactt
gataaagcac agaaaccaat tattatgcct 1140ttaaaacaat atcaacaaaa aattcaaaat
catgaatctt tt 1182101080DNABacillus thuringiensis
10atgcattcta ttaaaaaata taaaaaaatt ctattagttg caccacttgc ttgtatgtta
60acaggtgcta ttttacctac agctactaca gttcatgcac aagaaatcaa gggaccgggg
120gtaatgaaac ctgacgttcc gtggaatcaa gaacattata cgaaagaaaa tttagcatgg
180cgtgcagcag atagactttc ctatgctgcg gatagaattc ctagtttacg tgagaaattt
240aaattaaaac caaacgaaca tttttattgt agcaatgaca cgaggtacta tatggaagaa
300accttattaa aaaacttgca attatcagct gaaggtccaa taaatgttac accacatgta
360gatagttata ctgatttagg acaaacaaat ttattaacat ataacaatga tgatggaatt
420gtagagcaaa aggcttctac accagaaact accattaaag aatcagaaac atcttcttat
480tctaataaag aaggagttac attgggagca gaggtggaat ctaaagtaac attcaatata
540ccatttatag tagctggaga aacaaaggta atagcgaaat ccgaattttc ttatgaacat
600gatgatactc aaactaagac ccatgaaaaa gaggtaacat ttaaatcaca agagatcgtt
660gctgctccag agggaacaac tacttattat ggttcaatca aaactgccaa tttttctgga
720tcgttccaaa gtgatgctgt agtaggtggt ggtgtaacgt taacccttcc tataggagta
780atggataagg atggtgggca gaaaaaaact catacggaaa cagctacttt aactgcagaa
840gatatgtatg agattttcaa agcaccaatg ccttgggaca tgaataaatt accaccatat
900ctaaaattag atgattctgg caaaagggtg ctactggcag aaaaagcaac ctttgatata
960aagggacaag gtggttttta tacagaaata caggcaaaat ttgttccaaa agacaaaaat
1020aagaaaacac aaattatgcc atacgcagag tatgtacaga aagtaaaaca gaatgctctt
1080112028DNABacillus thuringiensis 11gtgaaaagta tgaattcata tcaaaataaa
aatgaatatg aaatattgga tgcttcacaa 60aataactcta ctatgtctac tcgttatcca
aggtatccac tagcaaagga tccacaagct 120tctatgcaga ctacgaatta taaagattgg
ctaaatctat gcgatactcc aaatatggaa 180aatccagagt ttcaatcagt aggaagaagc
gcactttcta ttctcattaa ccttagctct 240agaatattat ccttattagg tattcctttc
gcagcacaaa tcgggcaact ctggagctat 300acactcaacc tactatggcc tgtggcaaat
aatgctactc aatgggaaat ttttatgcgc 360accatagaag aattaattaa tgcacgcata
gagacttcgg taagaaatag agcccttgca 420gagctggcag gcttaggaaa catattagag
gactataagg tggttttaca acgatggaat 480ctaaatccta ctaatccaac attgcaacgc
gatgtggtac gccaatttga aatcgttcat 540gccttttttc gctttcaaat gccggtcttt
gctgtagatg gttttgaagt accattattg 600ccagtatatg cttcggcagc taatcttcat
ttgcttttac taagggatgt tgtaattaac 660ggagctcgtt ggggcttgga atctgatgta
attaacgatt atcatgacct tcaattacgt 720cttacatcaa catatgtaga ccattgcgta
acttggtaca acactggatt aaacaggtta 780attggcacaa atgctagaca atgggtaact
tacaatcagt tccgtaggga gatgactata 840tccgtactag atattatttc attattttct
aactatgatg ttcgtagata cccaacaaaa 900acacagagcg agctaacaag gatgatttat
acagatccaa taggtaccga agggaatcaa 960tttattcctg ggtgggtaga taatgcacct
tctttctcgg ttatagagaa tagtgtagtt 1020cgaagcccag gagctttcac ttttctggaa
agggttggta ttttcacagg gttcttacat 1080ggatggagta gccggtctga gttttggtca
gcccatagat tattttctag accggttttg 1140ggttggatat gggagagtgt tatttttggc
aatccccaaa ataatattgg gtatcaagaa 1200gtggatttta cgaattttga tgtatttagc
attaattcta gggccacttc tcatatgttc 1260ccaaatggca gtgctagatt atttggagtt
ccacgagtta catttgattt atcgaatgta 1320actaataata atctagcaca aagaacttat
aacagaccat ttacttttgg cggccaggat 1380atagtgtcga gattacctgg cgaaacaaca
gagataccga atagtagtaa ctttagtcac 1440agactagccc atatttcatc ttttccagta
ggtaacaatg gatcagtcct ctcatatggg 1500tggacacacc gtaatgtgaa tcgtcataat
agactgaatc cgaacagtat tacacagatt 1560ccagctataa agtttgctag tggttctgca
cggagaggtc ctgggcatac aggtggagat 1620cttgcaattg ctcaacaaca cagtggttat
cagctgttta tgcaatcgcc ttcagcacaa 1680aggtaccgtc tccgtttgcg ttatgccggt
atttcgggag gtagtatttc tgtttcgcat 1740cgggacgaaa ataatcaaaa catccttcat
agtgctacat tcaatgttag ggctacatca 1800ggtcagctaa gatacgccga tttcatttat
acagacctag aggagaacac aacgttgttt 1860gaaactcgaa atggagtgaa tctatataga
ctaatgattt ttgtttcaag tggctctata 1920ttaattgacc gaattgagta tatccctgaa
aatacaacaa ctatagaata tgaggaagaa 1980cgaaatctag aaaaagaaaa gaaagcggtg
gacgatttgt ttaccaat 2028121701DNABacillus thuringiensis
12gtgaataata tgtataccaa taatatgaaa actacattaa aacttgagac gacagattat
60gaaatagatc aagcggcaat ttcaatagaa tgtatgtcag atgaacaaga tctacaggaa
120aaaatgatgt tatgggatga agtaaaactt gcaaaacaac tcagtcaatc tcgtaattta
180ctctacaatg gtgattttga agattcatcc aacggctgga aaacaagtaa taatattacg
240attcaattgg agaatcctat tttaaaaggg aaatacctca atatgcctgg agcacgagac
300atatatggaa ccatatttcc aacatatgtt tatcaaaaaa tagatgaatc taaattaaaa
360cccaatacac gttatcgagt aagaggtttt gtgggaagta gtaaagatct aaaattagtg
420gtaacacgtt atgagaaaga aattgatgct agtatggatg ttccaaatga tttgtcctat
480atgcagccta gcccttcatg tggggattat ggctgtgact catcatccca gccaatgatg
540aatcaaggat atcctacacc atatacagac gactatgctt ccgatatgta tgcatgctcg
600tcaaacctag gtaaaaaaca tgtgaagtgt cacgatcgtc atccatttga ttttcatatt
660gacaccggag aattagatac aaatacaaac ttaggtattt gtatcttatt taaaatttcc
720aatccagatg gatatgctac attaggaaat ctagaagtaa ttgaagaagg accactaaca
780agcgaagcgt tagcacatgt gaatcaaaag gaaaagaaat ggaatcaaca aatggagaaa
840aagcgatcgg aaacacaaca agcctatgat ccggcaaaac aagcagtaga tgcattattc
900acaaattcac aaggagaaga gttacactat catattactt tagatcatat tcagaacgcc
960aatcagttgg tacagtcgat tccttatgta caccatgctt ggttaccgga tgccccagga
1020atgaactatg atttatataa caatttaaag gtacgtatag aacaagcacg ttatttatac
1080gatgcacgaa atgtcataac aaatggcgac tttgcacagg ggctaacggg gtggcacgca
1140acaggtaaag tagacgtaca acaaatggat ggagcttctg tattagttct atcaaactgg
1200agtgcggggg tatctcagaa tctgcatgcc caagatcatc atggatatat gttacgtgtg
1260attgccaaaa aagaaggtcc tggaaaaggc tatgtaacga tgatggattg taatggacat
1320caggaaacac tgaagttcac ttcttgtgaa gaggggtata tgacaaaaac agtagaggta
1380ttcccagaaa gtgatcgtgt acggattgaa ataggagaaa ccgaaggtac attttatata
1440gatagcatcg agttgctttg tatgcaagga tatgctagca ataatacccc acacacaggt
1500aatatgtatg agcaaagtta taatggaatt tataatcaga atacgagcga tctgtatcac
1560caagggtata caaacaacta taaccaagaa tctagtagta tgtataatca aaattatact
1620aacaatgatg accagcattc cggttgcaca tgtaaccaag ggcataattc tggctgtaca
1680tgtaatcaag gatataaccg t
1701131920DNABacillus thuringiensis 13atggaagaat tagagttaaa aagaacaaac
acactatctt ctgaggatgt gaatatttta 60caaattgaaa atttagtaaa agaatatgtt
aagcaaacat atggtaattc agctgaaatc 120aaaaaacttt cattagatgg gttagatgtt
ttgtataatt tagatattcc ttctatttta 180aagggtactt cttcttcttc tgctattaaa
gttggtactg acaatttgaa taacccaaca 240gatacagcga aaaccattaa acttccagtt
aaaaatgtac gaaaaaaaga atttaaggtt 300aaacctattc aagctttaaa ttttgaaaat
ggtgcaacaa tcactaaaaa aagtataact 360tcaattccta gtattaatgc tacttttata
gctttggctg aacagaattt tcaaaatgca 420cattttcata tagtaaacga tagtcaatct
tatgaaaatg aaatacccat ctatgtacct 480ccacattcaa aagttgaaat aacatactac
gtgaaagaaa tccaatttga tgcaattatt 540cagtccactg ccacaatagg tggatccata
agttttgaat atattgttca tgataatggc 600catgaaggaa tagattttct tactattttc
gaattagtaa atagccttaa tctaaatgac 660tttgaaattc aggaggcatc tgatgtacat
ggtaaggtag tttataaagg gaaatcccaa 720tttcagggaa ctgtaggttt aaatttattc
atgcaaatca aaggaacgcc attggatgaa 780agtaaaaata actatgaatt taccaaagta
ctatcagagg acgttgaaat gagtctatct 840ccgtcggagg gagaatatta tattgatttc
gggtcatctc ctaaattaac aaataaagaa 900gaagtaatag ttaaatttac aagagattac
cttttgtcaa atgaccgcaa aaacgcatac 960gtacaacaac taccacgtct tgaatatgga
gaagaagtta ctacattaaa atctattgat 1020actgctcatg aaaggaaaga aatcatagct
tctacaatta atacttttca aaatccttct 1080gatacagaga ttaccagaaa tacaataaaa
gagacattta gtacaacgga tacaattact 1140accactgcta caactgataa gtttcttgaa
ctgggaggaa gtatagaaac ctctgcaaaa 1200ggaaaagttc cactagtcgc agaagcatct
ataaaagtta cacagagtat aaaaggtggt 1260tggaaatggg taagtacaaa aactaataca
agaacaaacg tccatacaat tgaaatacca 1320tcgcaatcta ttaaaatacc tccacacaaa
atgtggaaat accagtatat tctaacaaaa 1380ttcgaatcaa gtggttattt aagctcagct
tgggaaataa atactaaaga atctatgtca 1440gctccggagg ttcacatagg ttactataat
aaagatcttc agaatccaag aaatataacg 1500gggctatcgg caaatgttga atctggaaat
gtagttggac gtgtatttga attcaataaa 1560ttccagccag gaggacttca ttacaaaata
ctgaatagtg aaaatatatt aaatgcaact 1620ccttatcaat tttttaaaga attagccaaa
cgtgttaatc aatacccatt aatacaaaat 1680aatcctcgct atagacgatt aggaattctt
ttgggatttg gaaaggacat atctcaaatc 1740acttgggaac ctcagataca ctataatgaa
catgtgttat ttgatgctga agaactatta 1800aatgttttac gttttgatga tattgccaat
aaagtatatg cacttgatgg aggcacacct 1860tttacagttg ctgtaggtca tgagctacta
ccaaaagagt caatagaacc attaaataat 1920142898DNABacillus thuringiensis
14gtgatgaata tgagtaatac cttagcacct tataatgttt tgagaagtat ggatatgccg
60aacatatcag gaaccaagtg ggataaagga atgtttatca atgcacttga taatacttct
120tttcttttag agcttataga aaaaggaatt aatgatgatg atgatgtgtt aggtctgtta
180agttttattg gattaacagc cttagaggca attccaattg tgggtggagt tatgtccaaa
240cttgtttcta tgatattttt ccctacaaaa tcaagcatta atttccagaa gatatgggag
300caattagaaa aagctattga acaaatagtt gacaaaaaaa taactgaagc tatgatgtct
360cagctaatgc aagaaatagc cggtttagcc gatgtattag aagaatatag gaatgcttat
420gatttatata atggtaaaaa attatttaat ataccagata agatgacacc tggggattat
480ctgatcaatg tatttactac tgcaaatttg caattcattc agagaatacc gacatttcag
540aactctatat atgatgtagt gtttcttcca ttctttgttc acgctgctga aatgcatatt
600cttctgatta gggatgcagc aatacatggt caagaatggg ggatggatga aactgtacac
660caaaaattta aaagggattt aaaaacttta attaataaat attctagtta tttattagct
720acatataaaa aaggattaaa agaagcatcc gaaaaaaaac ttgaaaataa tgattttcca
780acatctaaca accaacatca ttatattaat acagttagat ggaatgtaat caatcaatat
840aaaagaggga tggctttaac tgtttttgat tttgcttaca aatggaagta ttaccaagaa
900gtttatcaaa ataatataac gttaaatcct gctagaacaa tttattcaga tattgcaggt
960tcggtatatc cttatgaaaa aactacaaat gaaattgata atattatcaa agagcagaat
1020cttaaatatc gcggactctt aaaagaactg ctaattaatc atggggatag aatcgatagt
1080attcaaagta agtatataag gaacaatgaa ataattgata gtaacagaac tgggggggct
1140ggtggaaggg caaccttttt cgatttaaaa tctccaataa ataacccttt catacaagta
1200aatatgtggt ctgaattagt accattctct ttaggattca aatattataa tggggaggag
1260tcaaaactta tatggggagg agggacccct gggaaacata agtttggttc ttatcattat
1320gtagggaata aagtgtcttc tattatagga tttggtaaaa atggaaccgg tggattcaac
1380tctttagatg caatggtagt tggttttaaa cgagatgatt atatacctga aaatagattt
1440gttggtgtaa acaaaaatgg tgaacctgta actaaagtaa tagatgcaga gaatttctac
1500caagagaagt ttcaatcaaa tataaaaatg atagatgagc ctatgtttgg agaggcggtt
1560ttacaattcg aaaattattc taataatctt aataaggata gttatgtgac atatcaaatt
1620gatgcaaaga tagagggtac ttacgaatta catgtaatta taggtgcaaa aaaacaaaaa
1680gataaaatag cttttaaaat ggctcttaat gaaaaacagc cagaaaagtt tataactgaa
1740ccctttaatg ccggtgatat ttgggaagga atatcattga gcgaaggctt agtttataag
1800agaatattat taggaaattt ccaacttaag aaaggtatga atcgtattac tattcacaat
1860ggggtcctcc aaacatcagc aaatataaaa acatggaatt tagctaaact agagttaaca
1920ctcacttctg atagcttaaa agaccctgat attacaactt tatatgataa agataattat
1980tcaggaacaa aaaagtttat tttcgagaat acgagtcgtt taaaagactt taatgataaa
2040acatcatcaa taaaagtcga gtctcattta gctggcatta ggatttatca agattacaat
2100tataaaggta agtctatgga ccttgtaggt ggagaaaaaa taagtttaaa aaatcattca
2160tttaataaca gagcttcttc agttaaattt gctaatatcg ttttatataa ccaagataac
2220tatcagggtt caagaaaact agtttttgaa gatattcctg atttaggaaa acaaagcttt
2280aacgataaaa cctcttcaat tgttgttagt tctaatgtat ctggtgctag actatatgaa
2340catgcttatt ataaaggtaa gtatgtggat gttgttggtg gacagaaact taatttaaaa
2400aatcatgtat taaataaaaa gatttcatcc attaaatttt ttaaagaggg tgaagtactt
2460aatggtgtat atcaaattat tactgcaata aataacacga gtgtaataga taaacatcta
2520gaaaattcta acgttcattt atgggagaat gcagaaaaca aaaatcaaaa atggcgaatt
2580gagtatgatg tggctaaaaa agcttatcaa attaagaata tgttggatga gaagttagta
2640ttatcaacgc atgagttatt tccaattttc tcagcactat attgtttacc taacaaaggt
2700tatgtttcgc aatactggat ttttgagtat gtaggaaatg gttattatat tattaaaaat
2760aaagcgtatc ctgattgggt attagatgta gatggtttga attctgataa tggtacttta
2820attaaattac attcacagca tgatttaact gatccactta ttaatgcgca aaaatttaaa
2880cttaaggata taaataat
2898151725DNABacillus thuringiensis 15atgaatggaa atggaagaca tgatggatgg
aatcagaatc aacacataga aaatggacag 60atgaacccaa atcatagtgg atcttgtaag
tgcgggtgtc aacaaaataa taatggatcg 120tatccttcga atgagtataa ttcaaataat
aatggatcgt atccttcgaa tgagtataat 180tcaaataata atggatcgta tccttcgaat
gagtataatt caaataataa tggatcgtat 240ccttcgaatg agtataattc aaataataat
ggatcgtatc cttcgaatga gtataattca 300aataataatg gatcgtatcc ttcgaatgag
tataattcaa ataataatgg atcgtatcct 360tcgaatgagt ataattcaaa taataatgga
tcgtatcctt cgaatgagta taattcaaat 420aataatggat cgtattcttc gaatgagtat
aattcaaata ataatggatc gtatccttcg 480aatgagtata attcaaataa taatggatcg
tatccttcga atgagtataa ttcaaataat 540aatggatcgt atccttcgaa tgagtatgta
ggaggatata gtatacaaga tggtttacct 600caagaaagta aacagtttca aaaaatttca
aatatgaata ctagagataa tcatcgtgtt 660ttagacgcac aagatactta ttttggtcaa
ttgattgata atcgtgtagg tgacacatgt 720aaatatgtag agcataaaaa ttcagtaatt
tatgaactta gtaggcaacc tgtatatact 780cctgattccc aatatttcat tttttatcaa
atggataatg ggaattttat aattgcgaat 840aaagaaaata gtcgagtttt agaagttata
tttagttcag taaatggatt tgtaacaata 900tcaaatgagt ttaatgcaac ttcagatcaa
cgttttaagg ttgttagatc aaagaatgat 960acattccgct tagtaacaga aggaaataaa
acattaaata tatgtggtca ttcatttcaa 1020tataatacta aaattacagc tgtaaatgcg
gatattgatg gtgataatta tttatttcaa 1080aaatctatgg ataaggatac aagggattta
tatttcggaa caatatctaa taaaaatcca 1140gaaatattaa atgacccaag aaatttaaaa
agtttagatg atcttggtga tgagccgaga 1200gcatttaaag gagccgcatt acttcctgct
ctatttgtaa atgaccctag atattcagtg 1260catcgaagag tatcaaatag tccatattat
tacttagagt atacacagta ttggcataga 1320atatggactg atgttttgcc tattgatggt
tatggcgcat ggatagaaat gataggggta 1380acaaatgata cacaagtaaa tatgaaaaac
ataatgaata ttacaataac tggaaaagat 1440ttaggtgtag atttgggtat agatttggga
ttaagatttg gtgataagtc atttcttttt 1500gaacaaaaaa tcctttcagg attatctata
cggaaaactg attatccaaa tctcggaata 1560gatgaaagag caatgtatca aagaaacaat
agtaatttaa aaaccagatt tgtaagatac 1620gtgaaaaaac atgaatttgt attaagagat
ttgaatggga gtaaggtagc tgaaccatgg 1680attatcacgg aagatagatc gattacgaaa
gaatactctt caaat 1725161677DNABacillus thuringiensis
16atgaagtaca aaaatcgaac acgtgcaaaa tgcaaataca aacaagcgct tcttgtaaca
60gtagcaacaa tgacactagg agtaagtaca ttaggaagca atgcctcagc attcgctgat
120gaaaaagaaa agaatgttat tcagcaaaaa agtcctggca cttattatga agatgcgcaa
180aaaaatctgg gctccctagc acgatttgat acatgggcac aagatcttgg aaaaacaaca
240ggtgcaggga attataaaac tacacttggt atggcagaaa agctactacc aacaatttat
300aatgatttaa atagtggaaa tttcaacaat actgcaaggt ccattacgat gttatctaca
360gcgttgattc catatggagg agcattcatt tctccgataa ttgggatact ttggccagaa
420aacgggccaa atataaaaga aatgctgcag gaaatggaaa acaaacttgt tggtataatg
480gatgaaaaaa ttgaagccaa agatttagat gatcttgagg ctgcagtaaa aggattgatg
540gtaagtctaa aagaatttga aaactcattg aatggtaata taggtggtga atattattct
600gccctagctg atgtagactc acttaaccga ggtcgtataa cagccattca aaagggtttt
660aacgacctta ttagtgctac tagtaaaccg aagttcaaaa taacagaact tcctctatat
720acaattattg caactgctca cttgaatttc ttgaatactg tggaaaaaca gggaacttca
780cctaaaataa actacacaga agcagcctta aaagatcttc tacaaaatat gaagaagaat
840cacaaggatt atgcagatta tatagaaaaa acgtatacag aaggagaagc tagaatcaat
900agcaaactag aagataaaca aaaaatagaa caagatttag ctgccgtaaa ccaaaagcta
960tcggaaatgc ctcgtaaacc taagaatcac acccacgagg aagagaataa atttataatt
1020caaaaagaga agctttacgc gcaacaagac tctcttgaga aaaagttgtc tgaatacaat
1080gacttgatgt atcaaaagag tgatttttat agcaagacaa agggtagcga agcattccaa
1140atagcatcaa caggaaaaac gataccaact ccaagttggg ttaaaacaga aggaacgtgg
1200gtttgcgagg ctggtttttg gttttatatt gacgcaaaag ggcaaaagaa aagtgattgg
1260ttcaatgata agacacctga tggtaaggat agatggtatt accttagcac tgaaacacca
1320cgtctcgaca atgtaagggg gaatgcttat gttggaaaag gaacaatgct gactggttgg
1380ttccacgata cgcgtaaaga taagcagatc atcggtgtga atacgaagac tacttatgaa
1440tactggtatt acctcagccc agaaaaaaat cttaaaaact ctgccggaga actatttaag
1500cagggacaga tgatgacgaa atgggttgaa attaaggata caaagactgg tgaaccacac
1560tggtattatt tcaatcctga cgacggtagt atgacacatg ataaaaaagc ggtacaaatt
1620ggtgacaaaa aatatgattt tggttccaat ggtgtatgta caacgcctaa cggttac
1677173879DNABrevibacillus laterosporus 17atgaatcaga atcaaaataa
aaatgaaatg caaattatag aaccttcaag tgattctttt 60ctttatagtc acaacaatta
tccgtatgcc actgatccaa atacagtatt agaaggtagg 120aattataaag agtggctaaa
taagtgtaca gataattata cagacgcttt acagagtccc 180gaagctactg ctatttcaaa
aggagctgtt tctgctgcga tttctataag caccaaagtt 240cttggtttat taggtgttcc
atttgcagct caaatcgggc aactttggac cttcatatta 300aatgcgttat ggccttcaga
caatactcaa tgggaagagt tcatgagaca tgtagaagaa 360ctcataaacc aacgaatagc
cgattatgca agaaataagg cacttgcaga attaacgggt 420ttaggtaata atctagattt
atatatagag gctcttgatg attggaaacg aaatcctact 480agtcaagaag ctaaaacccg
tgtaatagat agattccgta tagtagatgg attatttgaa 540gcatatatac cttcatttgc
agtatcaggt tatcaagtac aattattaac ggtgtatgca 600gccgctgcaa atctccactt
acttttatta agagattcca ctatttacgg aattgattgg 660ggattaagtc aaactaatgt
taatgataat tacaatcgcc aaataagact caccgcaacg 720tatgcaaatc attgtacaac
ttggtatcaa actggtttag aaagattgag gggttccaat 780gcttccagtt gggtcactta
taataggttt cgaagggaaa tgacgttaac cgtattggat 840atttgttcct tattttcaaa
ttatgattat cgtagttacc cagcagaggt aaggggagag 900attacgagag aaatttatac
agacccagta ggtgtaggct gggtggatag tgcaccatca 960ttcggagaaa tagaaaatct
agcaattagg gcaccaagaa ccgttacttg gttaaattca 1020acaagaattt ttacagggag
attgcagggc tggagtggta ctaacaatta ttgggcagct 1080cacatgcaaa acttctcaga
aaccaattca ggaaatatac aatttgaagg tcctctctat 1140gggtcgacgg taggtactat
tcatcgtact gatgattacg atatggggaa tcgagatatt 1200tacaccatta cttcacaagc
tgttttaggc ctttgggcaa ctggtcaaag ggtgttgggg 1260gtcgcttcgg ctagatttac
tttaagaaat cttttcaata atcttacaca ggtgctggtg 1320tatgagaacc caataagttc
aacttttgga agttcaactt taactcatga attatctgga 1380gaaaactcag ataggccaac
ttctagcgac tatagtcata gactaacgag tatcacaggt 1440tttcgagctg gagctaatgg
aacggtccca gtgtttggtt ggacatctgc aactgttgat 1500cgtaacaata taattgagcg
aaacaaaata acacaattcc caggtgttaa gtcacacact 1560ctcaacaatt gtcaagtagt
taggggtact ggatttacag gaggagactg gttgagacca 1620aataataatg gtacatttag
actaactatt acttcattct ccagccaatc ttaccgaatt 1680cgcttacgtt atgctacttc
agtagggaat acttctttag ttatatcttc ttctgatgca 1740ggtatttctt ccacaacaat
tccgcttacc tcaacaataa catcactgcc acaaactgta 1800ccataccaag cttttagggt
tgtagattta cctattactt ttacaacacc tactacccaa 1860agaaattata cgtttgattt
ccgtctccaa aatccatcaa acgcaaatgt attcattgat 1920agatttgaat ttgttccaat
tgggggttct ttgtctgagt atgaaaccaa acatcagcta 1980gaaaaagcaa ggaaagcggt
gaacgatttg tttaccaatg aatcgaaaaa tgtgttaaaa 2040aaagacacga ccgattatga
tatagatcaa gctgcaaact tggtagaatg tgtatctgat 2100gaatgtgcaa atgctaaaat
gatcctatta gatgaagtaa aatatgcgaa acaacttagc 2160gaagcccgca atctacttct
aaatggtaat tttgaatacc aagatagaga tggggagaat 2220ccatggaaaa caagtcccaa
tgttaccatc caagagaata accccatttt taaaggccgc 2280tatctcagta tgtcaggtgc
gaacaatatc gaggcaacca atgagatatt tcccacttat 2340gtataccaaa aaattgatga
atccaaatta aaaccttata cccgttataa agttcgaggt 2400tttgttggaa atagtaaaga
tttagaatta ttggttacac ggtatgatga agaagtagat 2460gcgattttaa atgtaccaaa
tgatatacca catgctccgc cacctttctg cggtgaattt 2520gatcgatgca agccgcattc
ttatcctcct attaatccag aatgtcacca tgatgtaata 2580aataacattg aaatatcctc
tccttgccaa cacaataaga tggtagataa cgctgatata 2640tcttatcgcc atagccgatt
aagtaaaaaa catggcattt gtcatgaatc tcatcatttt 2700gaattccata ttgatacagg
gaaaatcgat ttggtcgaaa atttgggaat ttgggttgta 2760tttaaaatat gttccacaga
tggttacgca acattagata atttggaagt tattgaagag 2820ggtcctttag gagccgaatc
cttagaacgt gtgaaaagaa gagaaaagaa atggaaacat 2880cacatggaac acaaatgttc
agaaactaaa catgcatatc atgccgcaaa acaagcggtg 2940gtggcgttat tcaccaactc
taaatatgat agattaaagt tcgaaacaac catatccaat 3000attctttttg ctgattatct
cgtgcagtca attccgtatg tatataataa atggttacca 3060ggtgttccag gtatgaatta
cgatatctat acagaattaa aaaatctgtt tacgggagct 3120ttcaatctat atgatcagcg
aaatattata aaaaatggag actttaatcg tgggctcatg 3180cattggcatg cgacacctca
tgcaagagta gagcaaataa tagataatag gtctgtgcta 3240gtgcttccaa attatgctgc
caatgtttca caagaggttt gtttagaaca caatcgtggt 3300tatgtattac gtgtaacggc
gaaaaaagaa ggccctggaa ttggatatgt tacattcagt 3360gattgtgcaa atcatataga
aaagcttaca tttacttctt gcgattatgg tacaaacgta 3420gtgccatatg aacaatctaa
ttatcctaca gacggagtac catatggaca acatggttgt 3480aatatagacg gagtaccgta
tgaacaatcc ggttatcgta cagacggagt accgtatgaa 3540caatccggtt atcgtacaga
cggagtaccg tacgaacaat ctggtcatcg tacagatgga 3600gtaccgtacg aacaatctgg
ttatcgtaca gacggagtac catgcgaaca acatggttgt 3660catacagacg gactaccaca
catacaacat ggttgtcgta cagacggact accacacata 3720caacatggtt gtcgtacaga
cagatcaaga gatgaactac ttggttatgt gacaaaaacg 3780attgatgtat tccctaatac
agataaagta cgtatcgaca ttggagaaac cgaaggtact 3840tttaaagtag aaagtgtaga
actgatttgt atggaagag 3879183768DNABrevibacillus
laterosporus 18atgaatcaaa atcaaaatca gaatcaaaat aaaaatgaac tgcaaatcat
agaaccttca 60agcgattctt ttctttatag tcacaacaat tatccgtatg ccactgatcc
aaatacagta 120ttacaaggta ggaattacaa agagtggcta aacatgtgta caggtacaga
cgattcacga 180agtcccgaag ctgcttctac tgcaaaatca gctatttcag ttgcgattac
tataagcacc 240acaattcttg gcttactagg tgttccgttt gcatctcaaa tcggggcatt
ttataacttc 300gtattgaata cggtttggcc tcagggaaat aaccaatggg aagagttcat
gagacatgta 360gaagatctca taaacgaacg aatagctgat tatgcaagaa gtaaggcact
tgcagaatta 420gcgggtttag gtaataactt agatttatat agagaggctt ttgaagattg
gagacgaaat 480cctactagtc aacaagctaa aacccgtgta atagaaagat ttcgtatact
agatggtctt 540tttgaacaat atatgccatc atttgcagta caaggttttc aagtacaatt
attaacagtg 600tatgcatccg ctgcaaatat ccatttattt ttattaagag atagctctat
ttacggtttg 660gattggggat taagtcaaac taatgttaac gaaaattaca atcgccaaat
aaggcacgcc 720gcaacgtatg caaatcattg tacaacttgg tatcaaactg gtttacaaag
attgcaaggt 780accaatgcta ccagttgggt cgcttataat agatttagaa gggaaatgac
attaacagta 840ttagatatta gttcattatt ttcaaattat gattatcgta gttatccaac
agaggtaagg 900ggagagctta cgagagaaat ttatacggac ccagtaggta gaaactggca
gaatagtgca 960ccatcattcg ctcaaataga aaatctagca attagggcac caagaaccgt
tacttggtta 1020aattcaacaa gaatttctac agggaccttg cagggctgga gtggttctaa
cagatattgg 1080gcagctcaca tgcaaaactt ttcagaaacc aattcaggaa atatacgatt
tgacggtcct 1140ctatatgggt cgacggtagg tactattcat cgtactgatg attacgatat
ggggaatcga 1200gatatttaca ccattacttc agaagttgtt gcctcccttt gggcaactgg
tcaaactgtg 1260ttgggagtcg cttcggctag atttacttta agaaatcttt tcaataatct
tacacaggcg 1320ctggtgtatg agaacccaat aagttcaagt tttaataggt caactttaac
tcatgaatta 1380cctggagaaa actcagatag gccaacttct agcgactata gtcatagact
atcgagtatc 1440acaggttttc gagctggagc taatggaacg gtcccagtgt ttggttggac
atctgcaact 1500gttgatcgta acaatataat tgagcgaaac aaaataacac aattcccagg
tgttaagtca 1560cacactctca acaattgtca agtagttagg ggtactggat ttacaggagg
agactggttg 1620agaccaaata ataatggtac atttagacta actattactt cattctccag
tcaatcttac 1680cgaattcgct tacgttatgc tacttcagta gggaatactt ctttagttat
atcttcttct 1740gatggaggta tttcttccac aacaattccg cttacctcaa caataacatc
actgccacaa 1800actgtaccat accaggcttt tagggttgta gatttaccta ttacttttac
aacacctact 1860acccaaagaa attatacgtt tgatttccgt ctccaaaatc catcaaacgc
aaatgtattc 1920attgatagaa ttgaatttgt tccaattggg ggttctttgt ctgagtatga
aaccaaacat 1980cagctagaaa aagcaaggaa agcggtgaac gatttgttta ccaatgaatc
gaaaaatgtg 2040ttaaaaaaag acacgaccga ttatgatata gatcaagctg caaacttggt
agaatgtgta 2100tctgatgaat gtgcaaatgc taaaatgatc ctattagatg aagtaaaata
tgcgaaacaa 2160cttagcgaag cccgcaatct acttctaaat ggtaattttg ataacataga
tagagatggt 2220gagaatccat ggaaaacaag tcccaatgtt accatccaag agaataaccc
catttttaaa 2280ggccgctatc ttagtatgtc aggtgcgaac aatatcgagg caaccaatga
gatatttccc 2340acttatgcat accaaaaaat agatgaagca aaattaaaac cctatacgcg
ttataaagtt 2400cgagggtttg ttggaaatag taaagattta gagttgttgg ttacacggta
tgatgaagaa 2460gtagatgcaa ttttaaatgt accaaatgat ataccacatg ctccgccacc
tttttgcggt 2520gaatttgatc gatgcaaccc gcattcttat cctcctatga atccagaatg
tcaccatgat 2580gtaataaata acattgaaat atcctctcct tgccaacaca ataagatggt
agataacgct 2640gatatatctt atcgccatag ccataaaaaa catggcattt gtcatgaatc
tcatcatttc 2700gaattccata ttgatacagg gaaaatcgat ttggtcgaaa atttgggaat
ttgggttata 2760tttaaaatat gttccacaga tggttacgca acattagata atttggaagt
tattgaagag 2820cgtcctttag gagccgaatc attagaacgt gtgaaaagaa gagaaaagaa
atggaaacat 2880cacatggaac acaagtgttc agaaactaaa cttgcatatc atgctgcaaa
acaagcgctg 2940gtggggttat tcacaaacac tgaatatgat agattaaagt tcgaaacaac
catatccaat 3000attctttttg ctgattatct cgtgcagtca attccgtatg tatataataa
atggttacca 3060gatgttccag gtatgaattt cgagatctat acagaattaa aaaatctgta
tacgggagct 3120ttcaatttat atgatcagcg aaatattata aaaaatggag actttaatcg
cgggctcatg 3180cattggcatg cgacacctca tgcaagagta gagcaaatag ataataggtc
tgtgctggtg 3240cttccaaatt atgctgccaa tgtttcacaa gaggtttgtt tagaacacaa
tcgtggttat 3300gtattacgtg taacggcgaa aaaagaaggc cctggaattg gatatattac
attcagtgat 3360tgtgcaaata atatagaaaa gctgacattt acttcttgcg attatggtac
aaacgaagtg 3420ccgtatgagc aatctaatta tcctacagac ggagtttcat acggacacca
tggttgtaat 3480atagacagag taaggtacga agaatctggt tatcgtacag acggtgtacc
gtacgaacaa 3540tctggttatc gtgcagacgg agtatcgtac gaacaacatg gttgtcatac
cgacggagta 3600ccatacaaac aacatggttg tcgtacggac agatcaagag atgaacaact
tggttacgtg 3660acaaaaacga ttgatgtatt ccctgatact gataaagtac gtatcgacat
tggagaaacc 3720gaaggtacct ttaaagtaga aagtgtggaa ctgatttgta tggaagag
376819951DNABrevibacillus laterosporus 19atgaaaaaat ttgcaagttt
aattcttata agtgtgttcc ttttttcgag tacgcaattt 60gttcatgcgt catccacaga
tgttcaagaa agattacggg acttggcaag agaaaatgaa 120gctggaaccc ttaatgaagc
atggaatact aacttcaaac ccagtgatga acaacaattc 180tcttatagtc caactgaagg
tattgttttc ttaacaccac ctaaaaatgt tattggcgaa 240agaagaattt cacagtataa
agtaaataat gcatgggcta cattagaagg aagtccaacc 300gaagtatcgg ggacaccttt
atatgtggga aaaaacgtat tagataactc aaaaggaaca 360agcgatcaag agctgttaac
acccgagttt aactatacct atacggaaag cacttcaaat 420acaacaactc atggattaaa
attaggagtc aaaaccactg ctaccatgaa attcccgatt 480gctcagggta gcatggaagc
ttctactgaa tataactttc aagattcttc cactgatact 540acaactaaaa cagtatcata
taaaagccca tcacaaaaga ttaaagtacc agcaggtaaa 600acctttagag ttttagcata
cctaaatact ggatctattt caggtgaagc taacctttac 660gcaaatgttg ggggtatagc
ttggggggtt ttaccaggtt atcccaatgg cggaggagta 720aatataggtg ctgtacttac
caaatgccaa caaaaaggat ggggagattt cagaaacttt 780caacctagtg gaagagatgt
aatcgttaaa ggccaaggta ctttcaaatc taattatgga 840acggacttca ttttaaaaat
tgaagacatc acagattcaa agttacgaaa caataacggg 900agtggaactg tcgttcaaga
gattaaagtt ccactaatta gaactgaaat a 951203462DNABacillus
thuringiensis 20gtgaattttt tatttttagt taattatgaa aaaaataagt ttaaatataa
tatacaagga 60gacttgaata tgaatcaaaa aaactatgat attataggtt cttcgacaaa
cggcacaacg 120aaattacctg aagattataa cattataatt agtcctgatg cagccccaga
ggctgttact 180attgcgattt caattacagg agaagtactg tctctttttg gtgttccagg
tgcaacatta 240ggaagtactc ttcttaatac acttgttgat aaattatggc caaccaatac
aaatactgta 300tggggtacat ttacagagga aactgcaaaa cttataaatg aagtatataa
tccatcagat 360ccagtagtaa aagatgcaga tgctcgacta acatcgttac atgaatcgtt
aaaattatat 420caattagcct ttggaaattg gtttaaatca caagataatt caaaactcaa
agaagaggta 480cgacgccagt ttgatattac tcataataga tttgtaacta gtatgccttt
ttttaaggta 540tcggactacg aaattagatt gttaacaaat tatgctcaag ctgctaatct
tcatttaact 600tttttaagag atgcgtccat ttatgggctt gattggggtt tcagtgacga
gcatagtaat 660gatttgtatg aacaacaaaa gaatcgtaca ggagaataca cagatcattg
tgtaaagtgg 720tataacgcag gattagaaaa attaaaagga aatttaactg gggaaaattg
gtacacttat 780aatagatttc gtagagaaat gacgttaatg gtgttagacg tagttgcatt
atttccaaac 840tatgatacac gaatgtaccc gatcgcaacg tcatcagaac ttacaagaat
gatttataca 900gatccaatcg cttatacaca aagcgatcca tggtacaaga taacatctct
ttctttttcg 960aatattgaaa acagcgcgat tccaagtcct tctttcttca ggtggctaaa
atccgtttca 1020attaatagcc agtggtgggg cagtggtcct aatcaaacct actattgggt
tggacatgaa 1080ttggtatatt ctaattcaaa ttataatcaa tcacttaagg ttaaatacgg
ggatcccaat 1140tcttatattg agccccctga ttctttcagt ttttcttcta cggatgttta
cagaaccatc 1200tctgtcgtta gaaattcaat tagtaattat atagtaagtg aagttcaatt
caattcaatt 1260agtaatacaa atcaaattag tgaagaaatt tataaacatc aatcaaattg
gaatagaaga 1320gaaaccaaag attcaattac tgaactatcc ttagctgcta atcccccaac
aacatttgga 1380aacgtagcag aatacagtca tagattagca tatatttcag aggcatacca
aagtaacaac 1440ccatcaaaat acccagccta cattcctgta ttcggttgga cgcatacaag
cgtacgttac 1500gataataaaa ttttcccgga caaaatcact caaattccag ctgttaaaag
ttcctcagct 1560gaaggtggaa catggaaaaa tatagcgaaa ggtcctggat ttactggagg
cgatgtgaca 1620acagctgttt cgccagcatt tataacagat gtaataaaaa tacacgttac
tctagatcca 1680aattcacttt cacaaaaata tcgtgcacga cttcgctacg cttccaatgc
atatgtagca 1740gctacattgt atacaaattc aagtagtaat tataattttg aacttacaaa
aggtacaacg 1800gaacagttta caacatataa ttcataccag tatgtagata ttccaggttc
aatacaattt 1860aatactactt ctgatacagt gtctgtttat ttgcatatgg attcaacaac
taatgcaaac 1920gttcatgtag atagaattga attcattcca gtagatgaaa attacgataa
cagagtaaca 1980ctagaaaaag cacagaaagc cgtgaatgcc ttgtttacag cgggaagaca
tgcactccaa 2040acagatgtga cagattttaa agtagatcag gtttcaattt tagtggattg
tgtatcaggg 2100gaattatatc ccaatgagaa acgcgaacta ctcagtttag tcaaatacgc
aaaacgtttg 2160agttattccc gtaatttact cctagatcca acattcgatt ctattaattc
atctgaggag 2220aatggctggc acggaagtaa tggtattgca attggcaatg ggaactttgt
attcaaagga 2280aactatttaa ttttctcagg taccaatgat acacaatacc caacgtatct
ctatcaaaaa 2340attgatgaat ccaagctcaa agaatataca cgctataaac tgagaggatt
tatcgagagt 2400agtcaagatt tagaagcata tgtgattcgc tatgatgcaa aatatgaaac
attggatgta 2460tccaataatc tatacccaga tatttctcct gtaaatgcat gcggagaacc
caatcgttgt 2520gcggcactac catacctgga tgaaaatccg aggttagaat gtagttcgat
acaagatggc 2580attttatctg attcgcattc attttctctc aatatagata caggttctat
tgattccaat 2640gagaacgtag gaatttgggt gttgtttaaa atttccacac cggaagggta
tgcgaaattt 2700ggaaacctag aagtgattga agatggccca gtcattggag aagcattagc
ccgtgtgaaa 2760cgtcaagaaa cgaagtggag aaacaagttg acacaactgc gaacggaaac
acaagcgatt 2820tatacacgcg caaaacaagc cattgataat ttattcacaa atgcacagga
ctctcactta 2880aaaataggtg ctacattcgc gtcaattgtg gctgcgcgaa agattgtcca
atccatacgt 2940gaagcgtata tgccatggtt atctatcgtc ccaggtgtaa attaccctat
tttcacagag 3000ttgaatgaga gagtacagca agcatttcaa ttatatgatg tacggaatgt
cgtgcgtaat 3060ggccgattcc tgaatggagt atcggattgg attgtgacat ctgatgtaac
ggtacaagaa 3120gaaaatggga acaatgtatt agttctttcc aattgggatg cgcaagtatt
acaatgtctg 3180aagctctatc aagatcgcgg atatatcttg cgtgtaacgg cacgtaaaga
aggattggga 3240gaaggatata ttacaattac ggatgaagaa gggtatacag atcaattgac
atttggcaca 3300tgtgaggaga tagatgcatc taacacgttc gtatccacag gttatattac
aaaagaactg 3360gaattcttcc cagatacaga gaaagtgcgt atagaagtag gagaaacaga
aggaaccttc 3420cgggtagaaa gtgtagaatt attcttgatg gaagaacact gt
3462213618DNABacillus thuringiensis 21atgcggttaa aaaaattact
tgtatgtaat ataagaattg gaggaacaaa tatgaatttg 60ggaaactata atgaattcga
tatcatagat attactgaaa acaatcagac taaaacatca 120cgatataata atgtgaatag
acaagagaat ccatctaata tgattatttc aaatccttct 180tctaactatc ctctagcaaa
caatccaaat acacccttcc aaaatataaa ttataaagat 240tttttgaata tgaatgagga
gattgcaccg tatgcaagtt cgaaagatgt aatttttagt 300tcaatgaata tcattagaac
cttcatgggt tttgcaggac atgggactgc tggaggtatt 360gttgcattat ttacggaagt
attaagatta ctatggccta ataagcaaga tgagctttgg 420gaatcgttta tgaaagaagt
agagaaactt attgaacaag aaataacaga tgcggtagta 480agtaaagctt tggcagaatt
agagggttta agaaacgctt tgcaaggata tacagatgca 540ctggaagcat ggcaaaataa
tcgtagtgat aaacttaagc aattactagt gtatgataga 600tttgtttcta cagaaaattt
atttaaattt gcaatgccgt cttttagagt gggaggtttt 660gaagttccat tattaacagt
atatgcacaa gccgcaaatc ttcacttatt attattaaaa 720aattccgaat tattcggggc
agaatgggga atgcagcaat acgaaataga cctattttat 780aatgaacaaa aggattacgt
agtagaatat acagatcatt gtgttaaatg gtatactgaa 840gggttaaata ggttgaagaa
tgcaagcgga gtaaaaggta aggtatggga ggaatataat 900cgttttcgca gagaaatgac
gattatggtg ttagatcttc ttccattatt tccaatctat 960gatgtacgca catatcctac
ggaaacagta acagagttga caagacaaat tttcacagat 1020ccaataggtc ttagaggaat
taatgagtcg aaatatcctg attggtatgg agctgcaagt 1080gatagtttca gtcttataga
aaatagggca gtaccacaac ctagcttatt tcaatggtta 1140actgaattta aagtatatac
taaatatgtt gaaccgaatg ataagcttac aattttggct 1200ggacacagtg taactactca
atatactagc tattataaaa agagcacatt tacttatgga 1260gatacttcaa gtgctaattc
atctagaact tttgacctac ttgctaaaga tgtatatcag 1320gttgattctg tagctgcagc
aagtaaaagt gctacttggt atttggctgt tcctgaaatg 1380cgattatata gcattaatac
taacaatata ttatctgaag attatttttc tttgagtact 1440aatataccat ccagtaagat
gagacgtatg tattctagtg aggaattacc gataggaatc 1500tcggatacac ctatttatgg
agatcttgag gaatatagtc ataggttaag ttttatttct 1560gaaattatgc ataactctgg
aagtgtaaca ggttcaaata acatcaaagg aataattcca 1620gtattaggat ggacacatac
aagtgtatct cctgaaaatt atattcacag ggataaaatt 1680tcacaattat atgctgttaa
agcatacacc actagtaatg tttctgttgt aggaggacct 1740ggatttttag gaggaaatat
aattaaaggt aataatgatc ctgctagcta taccggaagc 1800gtgagctggg caattagatt
ggatggttca acagtaagtc gattccgtct tagaattccc 1860tatgctgctg aaacagatgg
cacattttct attactgttc gagacgattt aggccctttt 1920actataaaga aggactttat
agcaacaatg aaaccaggag atcctttatc atatggtaaa 1980tttgaatatt tagaatttga
acaaacaatg agtcttaata ataagcatgg tcaatttttc 2040gttcatacag aaaatttaaa
agatagaaat tctagtgtat attggaatag agttgaaatt 2100atcccggtgg atgaaaatta
cgataacaga gtaagattag aaaaagcaca gaaagccgtg 2160aatgctttgt ttacagcggg
aagacatgca ctccaaacaa atgtgacgga ttacaaagtg 2220gatcaggttt caatcttagt
ggattctgta tcaggggaat tatatccaaa tgagaaacgc 2280gaactacaaa gtttagttaa
atatgcaaaa cgtttgagct attcccgtaa tttacttcta 2340gatccaacat tcgattctat
taattcatct gaggagaatg gctggtacgg aagtaatggt 2400attgcaattg gaaatgggaa
ctttgtattc aaaggaaact atttaaattt ctcaggtacc 2460aatgatacac aatacccaac
gtatctctat caaaaaattg atgaatccaa gctcaaagaa 2520tatacacgct ataaactgag
aggatttatc gagagtagtc aagatttaga agcatatgtg 2580gttcgctatg atgcaaaaca
tgaaacattg gatgtatcca ataatctatt cccagatatt 2640tctcctgtga atgcatgcgg
agaacccaat cgttgtgcgg cactaccata cctggataaa 2700aatccgaggt tagaatgtag
tttgatacaa gatggtattt tatctgattc gcattcattt 2760tctctcaata tagatacagg
ttctattgat tccactgaga acgtaggaat ttgggtgttg 2820tttaaaattt ccacaccgga
agggtatgcg aaatttggaa acctagaagt gattgaatat 2880ggcccagtca ttggagaagc
attagcccgt gtgaaacgtc aagaaacgaa gtggagaaac 2940aagttgacac aactgcgaac
ggaaacacaa gcgatttata cacgagcaaa acaagccatt 3000gataatttat tcacaaatac
acaggactct tacctaaaaa taggtgctac attcgcgtca 3060attgtggctg cacgaaagat
tgtccaatcc atacgtgaag cgtatatgtc atggttatct 3120atcgtcccag gtgtaaatta
tcctattttc acagagttga atgagagagt acagcgagca 3180tttcaattat atgatgtacg
gaatgtcgtg cgtaatggcc gattcctgag tggagtatca 3240gattggattg tgacatctga
tgtaaaggta caagaagaaa atgggaacaa tgtattagtt 3300ctttccaatt gggatgcaca
agtattacaa tgtctgaagc tctatcaaga tcgcggatat 3360atcttgcgtg taacggcacg
taaggaagga ctcggagaag gatatattac aattacggat 3420gaagaagggc atacagatca
attgacattt ggcacatgtg aggaaataga tgcatctaac 3480acgttcgtat ccacaggtta
tattacaaaa gaactggaat tcttcccaga tacagagaaa 3540gtgcgcatag aaattggaga
aacagaggga atattcaagg tggaaagtgt agaattattt 3600ttgatggaag atctatgt
3618222316DNABacillus
thuringiensis 22atggtgaatg aaaatatgga tatgtataat aacaacggta gtatgaacgg
aaatccagat 60atgtacaata aaaacggaag tatgaacgga aatacggatg tgtataataa
caacggtagt 120atgaacggaa atccagatat gtacaataac aacggaagca tgaacggaaa
tacagatgtg 180tataataaca acggaagcat gaacggaaat ccagatgtgt ataataaaaa
cggaagcatg 240gacggaaatc cagatatgta caataacaac ggaagcatga acggaaatac
agatgtgtat 300aataaaaacg gaagcatgaa cggaaatcca gatatgtaca ataacaacgg
aagtatgaac 360ggaaatacgg atgtgtacaa taacaacgga agtatgaacg gaaatacgga
taatcaagtg 420ccggcttata acattctttc tgcggaaaac ccttctaata ttttagaaag
tgatactaga 480tgcacactaa atgttaaaaa tgtgcaagat gaggctatct gtacaggtag
taatttaacc 540aacgaaatag gtccacttgt tgttcccatt gcttttactc ctattattct
aacgcctgca 600cttattgaag taggtaaatg gttaggagtt caaattggta aatgggctct
aagtacagct 660ttaaaagaat taaaatcttt tctctttcca aattctgatc cccaaaggga
aatggagaaa 720ttacgcatag aattagaaaa ttcatttaat aagaaattaa cagaagataa
attgaatttt 780ttaactgccg cgtatactgg ttttaataat ttatctaatt cttttatttc
tgcaaccgag 840cgtgtaaaag cagcagaaat tacattagct acagctcctt ctcaagaaaa
tcaagatatt 900ttagatgaag ctagaacatt agcaagagac tattttgtga gtttacactc
acaaatgata 960gtgtggcttc cccagtttga aattagtgga tatgaagaaa tttccttacc
attatttact 1020cagatgtgca ctttacatct cactcatcta aaagatggag tattaatggg
gcagaattgg 1080gggctttcta cagatgatat taaacatttt aaaggtgaat tttacagatt
aagcaatgat 1140tacacttcta gagctttcga ttcatttcat agaggtttta atcgtttacg
aacacaacaa 1200ggaacagctg gagtcataaa atttagaaca gccatgaatg catatgcttt
tgacaatata 1260tataaatggt cattgttgcg ttatgagggt attaatccta ggataacaag
aagtttatgg 1320cattatattg gatataattc atctttagga tctaatgatt ttaatacact
atacaaactt 1380atggtgggta taccgcatga aagatttaga acagttgcaa taggatatcg
tgctaaaaca 1440ggtgaggatt ggaaagttac aggggctaaa tcaacttttt attctggtgg
tggtgaatgg 1500gttggaaacg tctccaaagc aacaagaatc cctgtttaca ctactaaaac
ggattggagg 1560caatttgaaa gaagaataca tggcagatta ggaactgagc aatatactag
atggcatctt 1620acaattcaag atacgaatat cattggtaat tcatatttaa ccggtttacc
ctttgatatt 1680tcatatcctg attattttat ccgaacaatt tcagcaaaac cagaagccta
ccctatttat 1740aaatcgctta gtctggggga taatccagga tacgtagtag acaatcctgg
aaataacctt 1800attataggtt tttctcctga taatttaaaa acatttatga ctgatggaaa
cagatatcat 1860tcaatagaat caggatatcc aacaaaccca tcttgtacta taccagcggt
actttataat 1920agtgtaagta acccattcca agcttatttt aatgatgaat taggtaatgg
ttcagatggg 1980agtataactt taatccgtcg aggtggtgca cattatcttg ttgattcaag
atctgcttct 2040tatgacagaa gctttcgtct tataattaga attcaggcag ggagttctgc
attcaaggta 2100acagtaagat caaggcacac atctgagagc tttgaattaa atttcacact
tctttcagat 2160caagatatta attattatta tgattatata tcccaacctt ttaatctaag
ctctacttac 2220tattatatag atgtagaacg tgttgttagt gatgacataa gagcgttaac
ttttaatcaa 2280atgattatag ttccaactac agaattccag atatta
231623867DNABacillus thuringiensis 23atggcaattt acgatatagc
agcagatttg ttcgatctta ccagatggta cgccgaacaa 60aattataatg caaatccaac
aacatttaga ggggctaaag tatatgatcg aatcgtatca 120gatgttcaat ctataccgga
aaaagtagat tttaatttga taccaggctt agcttatacg 180gtgaaaaatg aaatcgtaaa
tgatactaat acagaacaat ctatgagtac aaaactcatg 240catacattaa ttgaatcaaa
ttctgttaca accacaaaag gatataaaat tggaagtagt 300atcaaaaata cgtttagcgt
aaacattgag ggaagttttt ttgttggtgg tgggtctaca 360gagcattcca ttgaagtatc
agtaagtggg gaatataatc atagttcttc agaaactaaa 420acaaatacct cacagaaaac
atgggaatat aacagcccta ttcttgtccc agcaaaaaca 480aaagttacag caactttaga
tatttatgca ggaccagttg tagtcccagt aactttaaaa 540agtacagtta ctggaacggg
tattgtgaat aattttccta acgtattaac gagtttaagt 600tacattgata gaaacaataa
gttgtggaca gactctcttc caactgcttt gttatatgat 660tatcgaaatc agtggccagg
aagtcagtct atttatgttg ggaagaatgg gggaggtgta 720caggtagaag gtaaagctga
aatacaatta gaactaggtc tatactccat tgcaaccttc 780gactctcagc cactgtcagg
gaacacaaca ggtaaagaag cagtgtattc taaggctata 840ctccgagatg gatctattat
tgatatt 86724318DNABacillus
thuringiensis 24taaattcaat tatgattcgt aattaaaatg tgaaaattaa caatctccga
atttcatgtt 60aatttatggg tgtcaaataa aattatataa ggcctactta tattttcgag
attactatat 120gaggcctaaa atatttaaca aattgctatg tcttgcatat aaacaaattt
aagtatggtg 180aatttctcaa gatatatgta taaaaatgat aattatatcc tttgttgttt
tttgtgaata 240gatgaatgct aactattctt acctatgaaa aatagattaa tttgcctata
aagtattttt 300taaaaggagg aatgtatt
318253999DNABacillus thuringiensis 25ttggcacaat taaatgaaat
ttatccaagt tattacaatg ttttagcata tccccctcta 60attctcgacg ataagagcct
atatgatcag tatacagagt ggaagaaaaa aattgataag 120acttggaaac aatatgacaa
agacttttta ccaaagcctt taatggattt aggaaaatct 180ctggcagagg cctataaggg
tgatcctgat ggttaccttc atatcgcaaa cacagcaata 240agaatagctt tcttattgat
accaggaggg caaactgctg cttttggtgt aaatcttgtg 300ctgaataaag caatagggat
cttttatcct cctcaaaata aatctctatt tgatcaaatt 360aaggacgctg tatccaacct
ggtggatcaa aaattgatag accaagaaat ctctggagta 420ctgattaaac ttaacagcct
acagcagccc ctatcacgtt tcagcaattc catacagcga 480gctgttggaa aaccacagga
cttcgacgac caaactacat catccaacgc aattattctt 540gatgagacac aggactgcag
taaggacgat tcgtgttctt gttccaatac tcaacctcgt 600ccttctgatg cgcctctttg
taccccatgt atttgtcgta tgaaggaagt tcaacaaaca 660ttcaacaact caagtactga
tgttaaccga gcattaactg acatgaaaac aacgttaaag 720gacgtagtgg gtgcggatca
actgagaagc tacatgcaaa tatatttacc gttatatgtg 780acggctgcta caatggagtt
acaaatgtat aaaacctata ttgactttac acaaaaattc 840gatttcgatg taactggcac
gacaaaggaa catgtaaacg agttacgtca aaaaatcaaa 900acgcatagtg agtacatcat
gggcttattt aagaaatctt taccagaaat tagtaataat 960acaaaagaac aactaaatgc
atatattaaa tatacacgta atataacttt gaatgctctt 1020gatatggtaa gtacatggaa
atttttagac cctgttgact accctacgac tgctacattc 1080aatccgacta gaattatatt
taacgatctt gcaggacctg ttgagtgtct gaatagcact 1140caggacagca ataaactaca
ttttaatttc tttgatatga acggacagtc tatgcctaat 1200aatgatattt tcaattattt
ttatagaggt atgcaagtaa aaggtctgca gatccaaacc 1260tatactagct ctgacaccaa
aaatccacag cattttcctg taggattcct atcctcttat 1320tatggtagta acggtgattt
tccatttgac aagagagtag atcctaacaa atttacaggg 1380ggaagcaaat ccgtcaaact
gggagacgat gtatatgaga gccgttcagc tttaagtgta 1440ataaatgcag tgagtaatca
gctacaagtt tttctaaact atattgatac agaagatttg 1500tattttgacc aatctgtatc
ccctggtggc actgcatgtg ggtcaggtaa ttccacaatt 1560tggccagacc aaaaaattca
agctatatac cctatacaac ctgataattc tcaaacatat 1620ccaagttatt attcaacaag
taaaatagga tttgttacta cacttgtccc taatgatacc 1680actccatgga tcacttttac
cgataacggc aataacagca tttatacgtt ttctgcagaa 1740aatacacgaa ctcttacagg
ttcagcggga ccggttcgtg aatttataac tggttcagct 1800cctcttggac tgtctcctgg
aggcggtgca caatattcta ttaatactag tgatgctcct 1860agcggagatt atcaggttcg
cgttcatgta gccacacctg gctctggtgg gtcccttgct 1920atctcagttg atggaaaaac
gcaaacttta caactaccgg atacaaatgt gaacgataca 1980aaccatatag cgggattcgc
gggaacatat acgcttgctc ctgcaaccca agtagacgct 2040gcaaccctta aaccaaaggc
acctactgaa aatattttcc cagtgcgtca aacatcttct 2100ctacctgtga gcataaccaa
taattcttca acagttatta atatagaccg cattgaattt 2160gttcctgttt ctgctcctgc
tcctgatcct agtcctgact ctggtaagcc aatacacaaa 2220tcagtgccta agacagtgac
acaactaagc acaacgaaag agatttggtc atctactagc 2280gagtatgcta caactatatc
ttttacaggg aatgtttata acgatgcttc cattacattt 2340cagttattaa gttcaggtca
ggtagtgaaa gaatttccat ttaccggaaa cggggtcgcg 2400agtaaaccag gttttcatgg
cagttcaccg tcctgctatg acacgcctta tccattttcc 2460caaccggacc tatcagtgcc
taaatataat aaattgcagg tggtaatgaa gagcgatggt 2520tactcaaaac cttgtgatct
tggcgactca ttccctaata cttttgatgc ggaaatagac 2580ataaagttta atttgagtga
taccgcagac ttagcacaaa tcactgcaca agtgcaggga 2640ttattcacat cttcttcatc
tacagaatta tctccaaacg tttccggcta tcaaattgat 2700caaatcgcac tgaaagtgaa
tgcactatcc gatgaagtat tttgtaaaga aaaaatagta 2760ttacgcaaat tagtcaacaa
agccaagcaa ttcatgaaga cacgtaatct gctgataggc 2820ggggattttg aaatacttga
taaatgggca ttaggaacac aagctactat aaaagataat 2880tcatctttat ttaaagggaa
tcacttattt ttacaaccga ctaatggcat atcttcatct 2940tatgcttatc aaaaaataga
tgaatccaag ttaaaaccct acacacgcta taacgtttct 3000ggttttgtag cgcaaagtga
acacttagaa attgtcgttt ctcgctatgg gaaagaaatc 3060gataaaatat taaatgtccc
atatgaagaa gcattaccgg tttcttctgg gaatcagtcc 3120acttgttgca aaccatcttc
ttgctcatgt tcagcttgta ctggtggacc acatccacat 3180ttctttagct atagcattga
tgtcggtaag ctatatccag acttaaatcc aggaatagaa 3240tttggactgc gtcttgcaca
cccaagtgga tatgcaaaag tcggcaatct cgaaattgta 3300gaagaacgcc ctcttacaaa
cacagaaatt cgaaaaatcc aaagaaaaga agagaaatgg 3360aaaaaagcat gggatacaga
acgggcagaa attaatgcca tccttcagcc agtcattaac 3420cagatcaatg ctttctatac
aaatggagat tggaacggtt ctattcttcc tcatgtcaca 3480tatcaagacc tatataatat
cgtattacca gaattatcaa aattaagaca ttggtttatg 3540aaagatcgtc caggtgaaca
ctatactatc ctccaacaat tcaagcaagc tttagaacgc 3600gtattcaatc aattagaaga
aagaaactta atccacaatg gtagttttac aaatggatta 3660gcaaattggc tggtagacgg
agatacccaa ataactacct tagaaaatgg aaatctcgca 3720ttacaactct cagactggga
tgcaagcgca tcacaatcca ttgatatctc ggactttgat 3780gaagataaag aatatacagt
tcgcgtatat gcaaaaggaa aaggaaccat tagaactgta 3840aactgtgaaa atgagcccct
atcctttaat acaaacacat tcacaatcct agaacaacga 3900ttatatttcg acaacccatc
cgttctcctg cacatacaat cagaaggttc tgaattcgtc 3960ataggcagtg tggaactcat
tgaattgtca gacgacgaa 3999261134DNABacillus
thuringiensis 26ttgtattgta atacaatatt gcgaaaacgt tataaaaagt tagctacgat
tataccgctt 60acaagtatgt cagccgttgc gattgcacct gctacgtctt ttgcagttga
aacacaaaaa 120gcagatgttt catcacaaga agggccgatt caaggttacc agatggaaaa
tggaaaaatt 180actcctgtgt acaaaaataa acttacccag tttaatacgg cggatgatat
cgatcctggc 240cttccactac tcccagagaa tccatataat ccaattcctg atcatggaac
tgcatatgtt 300gaatcaactg atataggaga tactgtatat ttcaaaccat ttgaaccccc
taaaaataat 360gtattagagt taggtgactg tgatgataat acttatcagt ggtccgtatt
tgtagattca 420cagaaatata aaagtgtagg atactttgtt caaaaacaag ccgatggtca
aattagagtt 480ggatattata atccagaaga tttatctctg attacagatt caaaccatgc
tttcgcagga 540gtgccaggtt tcaaactgac agcagaagag aaagctgaga tgcaacgaga
tttaaatcga 600gaatatggcg atatatggga tggcacaagt aaactaaaac gagaaacaaa
ctataaactt 660ctgccaaatg cctcaggtct acaggatgac gcatcgggat ttggttataa
tcaaacatta 720acttcgggtg tatcaactac aaatatgttt ggaatagcga caacagttgg
gtggaaaatg 780gggataaaag tatcggttgt tcctcttgtt gcagacgtta cgtcagagat
tagtgcaagt 840ttaacagcta gttatcagca tactgtaaac gttacaaacc aaacgagttc
gcaagtgaaa 900tttgacgtat caagagtaga taaccctgac tataagtata atgactatgc
ggcagctgta 960tacaaaatat acacagacta tacattagaa ccgggtaaag gattatctcg
ttttttagca 1020aagcaagatc ttaaagatcc tgtgcgtaca gctgcattag caaatacgaa
ttatgcatat 1080gaaggttcaa aatactactt tacagtaaca cctggatcac acaagaaaat
tgtg 1134271149DNABacillus thuringiensis 27atggattttt taaattatta
taataaatta aaaaatgaat tggatgatgt aaattctaaa 60aaatattctt tagaatatac
atcagatgga ttaatggttc aacctactga tgatccatta 120aacacaatgc ctttgcctga
tagacctgta ttatctggaa acccaaatga ccctatccct 180tcagaaggaa caacacgtac
agatattcaa aaacaaaatc cacccttttt tacatttaaa 240gtagtagcta aattagctta
ttctggaaaa ggtgaaaatt gtcaaaaggc tcgtgcagca 300tcagtttatg gtgcagttct
tgaacttgaa aaagttaaac aattaccaga atattctaat 360gtatatttat actcggaaac
aggaataaaa acagatcgta gtaatataag atacaacacg 420gacggtataa tacaattttt
gaatcctagt tttataaaca cattttcttc aaatcctata 480aaatatgggg atacagtagg
ttatatttct tatccatatg atacattaaa atttccttct 540acaacacaat tagaacgttt
ggtgtacttt aatttactag atagtaatat tcttgataaa 600cacattggtt ttgattggtc
aaaaagtgtt acaaatggaa cagaagatac agaaatgtgg 660actcatagtt ccactgttgg
cgctgaatta aatcttaaag atatactaca gatcaatgca 720agttatgagc atacattttc
aacgtctcat atggagaaaa aagagaatac tgtatcaaaa 780actgctcatt ttaatagtcc
tttacctcct tataactatg ctacatgggt agcagctata 840tatcaattat ccattcgata
tcaacgtaca aatgcacaac cgatactaga tacgataaat 900gctgttaatt caggattaac
agcatctgaa acagatattt atctgaaggc attatacggt 960gctgggaaga atggtaagcc
tgctgtgggt gatccatcaa tattacacaa gttaagtaat 1020gtcatagaag atgcttatga
atatttatat tattcggata ctctttattt tactcaaact 1080ccttctggaa acagcccaac
accaaattct ccaaatcgca ttcaatttat tgccacagat 1140cctcaaagt
114928651PRTBacillus
thuringiensis 28Met Glu Ile Lys Ile Gly Ser Gly Gly Thr Tyr Met Asn Pro
Tyr Asn1 5 10 15
Asn Glu Ser Tyr Glu Ile Ile Asp Leu Asn Thr Ser Pro Tyr Pro Ser
20 25 30 Asn Arg Asn Asn Ser
Arg Tyr Pro Tyr Ala Asn Ala Cys Gly Phe Pro 35 40
45 Glu Asn Val Asp Trp Thr Ala Gly Ala Ser
Ala Met Ile Ile Val Ala 50 55 60
Gly Thr Leu Leu Ser Ala Ile Gly Ser Gly Gly Val Gly Ile Val
Ala65 70 75 80 Ala
Gly Ile Ile Ser Val Gly Thr Leu Phe Pro Phe Phe Trp Pro Gln
85 90 95 Asp Lys Pro Thr Ala Gln
Val Trp Lys Asp Phe Ile Lys Gln Gly Asp 100
105 110 Thr Ile Thr Asn Lys Thr Ile Ser Ala Ala
Val Glu Ser Leu Val Leu 115 120
125 Ala Glu Leu Asn Gly Leu Lys Ser Ile Leu Asp Val Tyr Thr
Asp Ala 130 135 140
Leu Glu Leu Trp Lys Lys Asp Lys Asn Asn Ile Val Asn Arg Asp Asn145
150 155 160 Val Lys Ser Ile Phe
Thr Asn Leu His Leu Gln Phe Val Ala Ala Met 165
170 175 Pro Lys Phe Ala Thr Asn Gly Tyr Glu Val
Ile Leu Leu Ser Thr Tyr 180 185
190 Thr Ala Ala Ala Leu Leu His Ile Thr Phe Leu His Glu Ala Leu
Gln 195 200 205 Tyr
Ala Asn Glu Trp Asn Leu Ala Arg Ser Glu Gly Thr Phe Tyr Arg 210
215 220 Gly Gln Leu Ile Gln Ala
Ile Glu Asn Tyr Ile Asn Tyr Cys Glu Lys225 230
235 240 Trp Tyr Arg Glu Gly Leu Glu Ile Leu Lys Asn
Ser Thr Trp Asp Ile 245 250
255 Tyr Ala Ala Tyr Gln Asn Glu Tyr Thr Leu Ser Ile Leu Asn Val Ile
260 265 270 Ser Ile Phe
Pro Arg Phe Asp Ile Arg Asn Phe Pro Thr Asn Ile Ala 275
280 285 Thr Arg Leu Glu Ser Thr Gln Lys
Leu Tyr Thr Thr Thr Pro Asn Met 290 295
300 Lys Ala Leu Lys Thr Asn Asn Ser Ile Asp Tyr Ile Lys
Asp Lys Leu305 310 315
320 Ile Pro Pro Leu Asp Leu Phe Lys Lys Leu Lys Ser Leu Thr Phe Tyr
325 330 335 Thr Phe Leu Asp
Ser Asn Asn Gln Tyr Asp His Leu Gln Gly Ile Val 340
345 350 Asn Asn Ser Tyr Tyr Thr Asn Ile Ser
Thr Asn Lys Ile Phe Ser Ser 355 360
365 Gly Thr Thr Glu Gly Ser Ser Tyr Gln Leu Gly Leu Ala Ser
Asp Gln 370 375 380
Val Ile Tyr Tyr Thr Asp Ile Phe His His Leu Asn Gln Ser Asn Phe385
390 395 400 Lys Asp Gly Ser Leu
Gly Ile Lys Ile Ile Asn Phe Asn Ile Ile Asn 405
410 415 Lys Tyr Asn Glu Val Ser Gln Lys Ser Tyr
Asp Ser Asn Ala Thr Ser 420 425
430 Asn Leu Ile Leu Glu Val Ile Leu Pro Phe Leu Lys Thr Thr Glu
Lys 435 440 445 Asp
Tyr Lys Tyr Ile Leu Ser Tyr Ile Thr Ile Thr Pro Gln Gln Ile 450
455 460 Val Gly Cys Leu Ser Pro
Ser Tyr Ile Tyr Gly Phe Ile Trp Thr His465 470
475 480 Ser Ser Val Asn Leu Asn Asn Thr Ile His Tyr
Thr Asn Lys Asn Asn 485 490
495 Phe Ser Gln Ile Thr Gln Ile Ser Ala Val Lys Ala Tyr Leu Lys Lys
500 505 510 Asp Arg Val
Ser Val Ile Glu Gly Pro Gly His Thr Gly Gly Asp Leu 515
520 525 Val Lys Phe Thr Gln Trp Asp Asp
Ser Ile Ser Thr His Tyr Gln Phe 530 535
540 Thr Ser Ser Gly Glu Tyr Lys Ile Arg Val Arg Tyr Ala
Ser Thr Ala545 550 555
560 Gln Val Asn Gln Thr Ser Gly Leu Ser Met Thr Ile Tyr His Lys Gly
565 570 575 Asn Pro Thr Glu
Thr Trp Asp Leu Asn Ile Asn Asn Lys Ser Asp Thr 580
585 590 Ile Leu Asn Leu Asn Glu Pro Lys Tyr
Asn His Phe Gln Tyr Thr Glu 595 600
605 Phe Pro Asn Lys Thr Leu Ile Ile Asn Lys Asp Pro Asn Ser
Pro Tyr 610 615 620
Leu Glu Leu Arg Ile Asp Leu Ser Tyr Lys Gly Asn Thr Ala Thr Thr625
630 635 640 Leu Ile Asp Lys Ile
Glu Phe Ile Pro Val Ser 645 650
29506PRTBacillus thuringiensis 29Met Asn Gly Asn Gly Lys His Asp Asn Trp
Asn His Asn Gln Gln Ile1 5 10
15 Ser Asn Val Gln Met Asn His Asn His Gly Arg Ser Tyr Asp Cys
Ser 20 25 30 Cys
Gln Gln Asn Gln Tyr Gly Tyr Glu Gln Gln Lys Gln Gln Tyr Glu 35
40 45 Gln Asn Asn Ser Gln Tyr
Met Gln Asn Asn Leu Gly Asn Glu Asn Arg 50 55
60 Asn Gly Leu Tyr Pro Tyr Gln Glu Asn Gln Tyr
Glu Gln Asn Lys Asn65 70 75
80 Tyr Tyr Ala Ser Asn Asn Leu Thr Tyr Asn Gln Ser Asp Leu Tyr Asn
85 90 95 Ser Asn Pro
Gln Asn Met Tyr Lys His Gln Thr Tyr Ser Asn Asp Phe 100
105 110 Tyr Cys Ser Pro Ser Tyr Thr Ala
Gly Glu Asn Asn Ile Leu Asp Leu 115 120
125 Leu Gly Thr Glu Ser Lys Gln Phe Gln Lys Ile Ser Asn
Ile Asn Thr 130 135 140
Lys Asp Leu His Arg Ser Ile Thr Ala Ser Asn Thr Gln Ile Gly Tyr145
150 155 160 Gln Ile Asp Thr Arg
Val Pro Gly Pro Cys Lys Gly Val Asp Tyr Gln 165
170 175 Asn Thr Val Thr Tyr Glu Gln Asn Ser Ile
Gly Gly Asp Ser Gln Tyr 180 185
190 Leu Ile Phe Tyr Lys Thr Asp Tyr Thr Asp Ala Phe Ile Ile Ala
Asn 195 200 205 Arg
Ala Asn Gly Arg Val Leu Glu Val Ile Pro Ser Ser Val Asn Gly 210
215 220 Phe Val Thr Ile Ser Asn
Met Phe Thr Tyr Asn Gln Asn Gln Leu Phe225 230
235 240 Ile Arg Thr Lys Ile Ser Asn Asn Asp Asn Ser
Asp Asp Val Pro Phe 245 250
255 Ser Leu Thr Thr Glu Asn Asn Gln Thr Leu Asn Ile Cys His His Glu
260 265 270 Phe Gln Tyr
Asn Thr Lys Ile Thr Ala Leu Asp Asn Ala Tyr Arg Leu 275
280 285 Asp Asp Lys Val Leu Phe Lys Pro
Thr Arg Asp Lys Ile Asn Ile Ser 290 295
300 Phe Pro Asn Met Val Val Asn Ala Lys Glu Lys Leu Pro
Glu Pro Glu305 310 315
320 Glu Leu Thr Asn Met Asp Lys Asn Thr Leu Phe Ile Pro Lys Val Ile
325 330 335 Ile Ser Lys Thr
Leu Ile Pro Gly Ile Ile Val Asn Asp Val Thr Leu 340
345 350 Leu Lys Glu Gln Gln Ile Ala Lys Ser
Pro Tyr Tyr Val Leu Glu Tyr 355 360
365 Val Gln Ser Trp Glu Glu Val Tyr Asn Glu Ile Val Pro Ala
Tyr Arg 370 375 380
Pro Ser Tyr Thr Trp Thr Ser Thr Asp Gly Ile Arg His Val Asn Leu385
390 395 400 Leu Asp Ile Lys Asn
Thr Ile Asn Ile Ser Ile Gly Gly Thr Ser Gln 405
410 415 Gly Trp Gly Leu Arg Phe Ser Asp Lys Ser
Asp Leu Phe Lys Asn Ile 420 425
430 Ile Thr Ser Ala Phe Ile Ile Lys Ser Thr Gln Ala Pro Asp Met
Gly 435 440 445 Phe
Ser Glu Asn Asp Ile Asp Gln Tyr Tyr Gly Lys Asn Ile Asp Ser 450
455 460 Arg Val Lys Ile Tyr Ile
Lys Thr His Asn Leu Ile Leu Arg Arg Leu465 470
475 480 Asp Gln Leu Asn Asn Ser Ile Ala Thr Trp Thr
Ile Phe Glu Asn Thr 485 490
495 Lys Pro Val Ile Arg Thr Phe Pro Ile Ser 500
505 30779PRTBacillus thuringiensis 30Met Lys Asp Lys Lys Tyr
Trp Lys Tyr Glu Gly Gly Thr Lys Met Asn1 5
10 15 Pro Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Val
Asn Asn Pro Gln Asn 20 25 30
Tyr Asn Thr Val Ser Asn Arg Tyr Pro Tyr Thr Asn Asp Pro Asn Val
35 40 45 Ala Ile Gln
Asn Thr Asn Tyr Lys Asp Trp Met Asn Gly Tyr Glu Glu 50
55 60 Ile Asn Pro Ser Ser Ile Ser Leu
Ile Leu Ala Ser Ile Gly Ile Leu65 70 75
80 Asn Gln Ala Ile Ala Leu Thr Gly Val Leu Gly Lys Thr
Pro Glu Ile 85 90 95
Ile Asn Ile Val Gln Glu Met Val Gly Leu Ile Ser Gly Ser Thr Gly
100 105 110 Asn Asp Leu Leu Val
His Thr Glu Gln Leu Ile Gln Gln Thr Leu Ala 115
120 125 Gln Gln Tyr Arg Asn Ala Ala Thr Gly
Ala Val Asn Ala Ile Ser Lys 130 135
140 Ser Tyr Asn Asp Tyr Leu Met Phe Phe Arg Gln Trp Glu
Arg Asn Arg145 150 155
160 Thr Ser Gln Asn Gly Leu Gln Val Glu Ser Ala Phe Asn Thr Val Asn
165 170 175 Thr Leu Cys Leu
Arg Thr Leu Thr Pro Gln Glu Ala Leu Ser Arg Arg 180
185 190 Gly Phe Glu Thr Leu Leu Leu Pro Asn
Tyr Ala Leu Ala Ala Asn Phe 195 200
205 His Leu Leu Leu Leu Arg Asp Ala Val Leu Tyr Arg Thr Gln
Trp Leu 210 215 220
Pro Asn Phe Ile Ser Thr Thr Asn Ala Asn Ile Glu Ile Leu Glu Arg225
230 235 240 Ser Ile Asn Gln Tyr
Arg Asn His Cys Asn His Trp Tyr Asn Asp Gly 245
250 255 Leu Asn Arg Phe Ala Arg Thr Ser Phe Asp
Asp Trp Val Arg Phe Asn 260 265
270 Ala Tyr Arg Arg Asp Met Thr Leu Ser Val Leu Asp Phe Val Thr
Val 275 280 285 Phe
Pro Thr Tyr Asn Pro Ile Asn Phe Pro Thr Pro Thr Asn Val Glu 290
295 300 Leu Thr Arg Ile Val Tyr
Thr Asp Pro Ile Ser Pro Pro Arg Gly Tyr305 310
315 320 Ala Arg Thr Gly Ser Pro Ser Phe Arg Gln Met
Glu Asp Leu Ile Ile 325 330
335 Ser Gly Ser Pro Ser Phe Leu Asn Gln Leu Ser Ile Phe Thr Thr Tyr
340 345 350 Tyr His Asp
Pro Arg Asn Val Asn Arg Asp Phe Trp Ala Gly Asn Arg 355
360 365 Asn Tyr Leu Ser Asn Gly Thr Ser
Arg Gln Ser Gly Ala Thr Thr Pro 370 375
380 Trp Arg Thr Asn Ile Pro Met Gln Asn Ile Asp Ile Phe
Arg Val Asn385 390 395
400 Leu Thr Thr His Asp Ile Asp Asp Ile Ser Arg Ser Tyr Gly Gly Val
405 410 415 His Arg Ser Asp
Phe Ile Gly Val Asn Thr Ile Asn Asn Gln Arg Thr 420
425 430 Thr Leu Phe Tyr His Gln Asn Val Asp
Thr Ser Arg Phe Leu Ile Arg 435 440
445 Asn Glu Thr Val Phe Leu Pro Gly Asp Ser Gly Leu Ala Pro
Asn Glu 450 455 460
Arg Asn Tyr Thr His Arg Leu Phe Gln Val Met Thr Thr Tyr Arg Thr465
470 475 480 Asn Pro Asn Ala Arg
Arg Ala Ala Phe Leu His Ala Trp Thr His Arg 485
490 495 Ser Leu Arg Arg Arg Asn Gly Phe Arg Thr
Asp Gln Ile Met Gln Ile 500 505
510 Pro Ala Val Lys Ser Ile Ser Asn Gly Gly Asp Arg Ala Val Ile
Ser 515 520 525 Tyr
Thr Gly Glu Asn Met Met Lys Leu Asp Asn Leu Thr Ala Ser Leu 530
535 540 Ser Tyr Lys Leu Thr Ala
Glu Asp Ser Glu Ala Ser Asn Thr Arg Phe545 550
555 560 Ile Val Arg Ile Arg Tyr Ala Ser Met Asn Asn
Asn Arg Leu Asn Leu 565 570
575 Ile Leu Asn Gly Thr Gln Ile Ala Ser Leu Asn Val Glu Gly Thr Met
580 585 590 Gln Asn Gly
Gly Ser Leu Thr Asn Leu Gln Ser Glu Asn Phe Lys Tyr 595
600 605 Ala Thr Phe Ser Gly Asn Phe Lys
Met Gly Ser Gln Ser Ile Val Gly 610 615
620 Ile Phe Lys Glu Ile Ser Asn Ala Asp Phe Ile Leu Asp
Lys Ile Glu625 630 635
640 Leu Ile Pro Ile His Phe Met Pro Leu Leu Glu Gln Lys Gln Ser Tyr
645 650 655 Asn Asn Tyr Asp
Gln Asn Met Asp Thr Thr Tyr Gln Pro Asn Tyr Asp 660
665 670 Thr Tyr Asn Gln Asn Ala Asn Gly Met
Tyr Asp Asp Thr Tyr Tyr Pro 675 680
685 Asn Asn Asn Asp Ser Tyr Asn Gln Asn Asn Thr Asp Met Tyr
Asp Ser 690 695 700
Gly Tyr Asn Asn Asn Gln Asn Thr Asn Tyr Asn Tyr Asp Gln Glu Tyr705
710 715 720 Asn Thr Tyr Asn Gln
Asn Met Glu Asn Thr Tyr Asp Gln Ser Tyr Glu 725
730 735 Asn Tyr Asn Pro Glu Thr Asn Asn Tyr Asn
Gln Tyr Pro Asn Asp Met 740 745
750 Tyr Asn Gln Glu Tyr Thr Asn Asp Tyr Asn Gln Asn Ser Gly Cys
Arg 755 760 765 Cys
Asn Gln Gly Tyr Asn Asn Asn Tyr Pro Lys 770 775
31322PRTBacillus thuringiensis 31Met Asn Phe Leu Tyr Asn Phe Val
Thr Leu Asp Met Leu Ile Leu Asn1 5 10
15 Arg Leu Glu Gly Ser Asp Phe Lys Met Lys Lys Lys Ala
Ile Val Cys 20 25 30
Gly Leu Leu Ala Ser Thr Leu Leu Gly Gly Gly Thr Phe Val Asp Ala
35 40 45 Val Ser Ala Ala
Glu Ile Gln Lys Thr Asn His Leu Asn Lys Tyr Asp 50 55
60 Ser Ala Gln Glu Lys Ala Leu Gln Asp
Ile Asn Gln Glu Ala Leu Gln65 70 75
80 Asp Ile Asp Gln Lys Val Asn Lys Met Ile Asp Ser Ile Pro
Pro Ile 85 90 95
Phe Gly Ser Lys Tyr Thr Arg Thr Asp Arg Tyr Gly Glu Ser Leu Thr
100 105 110 Tyr Ser Gly Ile Asn
Leu Lys Glu Asn Asn Ser Thr Asn Val Glu Pro 115
120 125 Met Tyr Phe Gly Ser Asn Thr Phe Tyr
Asn Asp Thr Glu Leu Glu Gln 130 135
140 Ser Tyr Asn Thr Thr Ser Phe Ser Glu Ala Val Thr Lys
Ser Thr Thr145 150 155
160 Thr Gln Thr Gln Asn Gly Phe Lys Ser Gly Val Thr Thr Gly Gly Lys
165 170 175 Val Gly Ile Pro
Phe Val Ala Glu Gly Glu Val Lys Ile Asn Leu Glu 180
185 190 Tyr Asn Phe Thr His Thr Asn Ser Asn
Thr Thr Ser Lys Thr Thr Thr 195 200
205 Leu Thr Ala Pro Pro Gln Pro Val Lys Val Pro Ala Gly Lys
Val Tyr 210 215 220
Lys Ala Asp Val Tyr Phe Glu Lys Lys Ser Thr Ser Gly Thr Val Glu225
230 235 240 Leu Tyr Gly Asp Leu
Leu Thr Gly Val Val Ala Glu Gly Arg Thr Ser 245
250 255 Phe Val Gly Asn Val Leu His Lys Ala Thr
Asp Thr Gln Gly Leu Ile 260 265
270 Gln Ser Pro Glu Asp Ser Asn Lys Val Arg Ala Val Gly Lys Gly
Thr 275 280 285 Phe
Thr Thr Glu His Gly Ser Asn Phe Ile Val Lys Thr Tyr Asp Val 290
295 300 Thr Ser Gly Gln Lys Ser
Ala Lys Leu Val Asp Thr Arg Val Ile Pro305 310
315 320 Ile Lys32323PRTBacillus thuringiensis 32Met
Asn Leu Arg Arg Ile Ser Met Arg Val Tyr Lys Lys Leu Ala Thr1
5 10 15 Leu Ala Pro Ile Ala Ala
Leu Ser Thr Ser Ile Leu Cys Ser Pro Ala 20 25
30 Met Thr Phe Ala Ala Glu Lys Glu Ser Thr Val
Lys Gln Thr Thr Gln 35 40 45
Gln Ser Ala Val Gln Gln Gly Arg Ile Ile Gln Gly Tyr Leu Ile Lys
50 55 60 Asn Gly Val
Lys Ile Pro Val Tyr Thr Gly Gly Leu Val Thr Asn Lys65 70
75 80 Ala Glu Gln Gly Ala Ala Ala Phe
Pro Gln Leu Ser Ser Asn Pro Asn 85 90
95 Asp Pro Ile Pro Gln Lys Gly Ser Ile Ser Ser Glu Asp
Gly Asn Ile 100 105 110
Gly Asp Ile Leu Tyr Phe Ser Lys Thr Pro Met Gly Asp Asn Val Tyr
115 120 125 Ile Lys Lys Leu
Glu Asn Asn Asn Ile Glu Ile Gly Lys Tyr Asn Arg 130
135 140 Gly Thr Leu Glu Leu Ser Lys Phe
Val Thr Val Asn Gly Asp Pro Gln145 150
155 160 Gly Pro Ile Met Leu Phe Asp Ala Thr Val Lys Arg
Glu Thr Ala Phe 165 170
175 Glu Lys Ile Gly Gly Ala Val Gln Pro Lys Ala Thr Gln Tyr Thr Phe
180 185 190 Ser Gln Ala
Val Thr Ser Gly Leu Ser Thr Ser Asp Ala Ile Gly Gly 195
200 205 Ser Leu Thr Leu Gly Tyr Lys Ile
Ser Leu Lys Glu Gly Gly Gly Val 210 215
220 Val Pro Ala Glu Ala Thr Gln Glu Phe Ser Thr Gln Leu
Ser Ala Thr225 230 235
240 Tyr Asn His Thr Ile Thr Val Thr Asn Gln Thr Thr Asn Thr Gln Thr
245 250 255 Gln Thr Phe Lys
Pro Ile Asp Ser Tyr Gly Gln Ser Thr Tyr Ala Ala 260
265 270 Ala Val Tyr Gln Leu Lys Ser His Tyr
Thr Val Ile Pro Gly Ala Gly 275 280
285 Leu Gln Lys Gly Leu Asn Ser Gly Tyr Val Leu Asp Gln Thr
Ala Phe 290 295 300
Ser Tyr Ser Asp Ser Asp Leu Tyr Leu Ala Val Thr Pro Gly Ala Gly305
310 315 320 Ser Asn
Val33252PRTBacillus thuringiensis 33Met Ile Thr Asn Gln Ala Ala Gln Ala
Ser Asp Ala Pro Tyr Pro Glu1 5 10
15 Leu Pro Ser Asn Pro Asn Asp Ala Ile Pro Asn Ala Gly Ala
Thr His 20 25 30
Ala Glu Asn Gly Ser Val Gly Ser Val Leu Tyr Phe Lys Gln Ile Asp 35
40 45 Leu Asn Asn Leu Gly
Ala Gly Ile Gly Asn Ser Gln Lys Asp Tyr Val 50 55
60 Tyr Val Glu Lys Lys Gly Asp Ser Gly Tyr
Glu Leu Gly Asn Tyr Asn65 70 75
80 Pro Leu Thr Leu Gln Arg Thr Lys Ile Lys Asp Tyr Asp Lys Ser
Ser 85 90 95 Glu
Leu Ala Glu Lys Met Asp Gly Tyr Phe Lys Ser Thr Ile Thr Arg
100 105 110 Asp Thr Phe Phe Ser
Lys Ile Gly Ser Gly Val Val Pro Lys Asn Ala 115
120 125 Ala Tyr Thr Phe Ser Gln Ala Val Thr
Ser Gly Leu Thr Thr Ser Asp 130 135
140 Ala Ile Gly Gly Ala Leu Thr Leu Gly Tyr Lys Val Ser
Val Thr Glu145 150 155
160 Gly Gly Gly Ile Phe Pro Ala Ala Ala Ser Glu Glu Phe Ser Ala Gln
165 170 175 Leu Thr Ala Thr
Tyr Asn His Thr Ile Thr Val Ser Ser Gln Val Thr 180
185 190 Asn Thr Gln Thr Leu Gly Ile Thr Lys
Ala Ala Asp Gly Tyr Gln Tyr 195 200
205 Asp Lys Tyr Val Gly Ala Val Tyr Gln Leu His Ser Lys Tyr
Thr Phe 210 215 220
Lys Pro Ser Asp Glu Leu Gln Phe Ala Met Asn Ser Pro Phe Gly Tyr225
230 235 240 Lys Val Ile Leu Asn
Gln Arg Ala Gln Ser Phe Gln 245 250
341441PRTBacillus thuringiensis 34Met Thr Thr Ile Asn Glu Leu Tyr Pro
Ala Val Pro Tyr Asn Val Leu1 5 10
15 Ala Tyr Ala Pro Pro Leu Asn Leu Ala Asp Ser Thr Pro Trp
Gly Gln 20 25 30
Ile Val Val Ala Asp Ala Ile Lys Glu Ala Trp Asp Asn Phe Gln Lys 35
40 45 Tyr Gly Val Leu Asp
Leu Thr Ala Ile Asn Gln Gly Phe Asp Asp Ala 50 55
60 Asn Thr Gly Ser Phe Ser Tyr Gln Ala Leu
Ile Gln Thr Val Leu Gly65 70 75
80 Ile Ile Gly Thr Ile Gly Met Thr Val Pro Val Ala Ala Pro Phe
Ala 85 90 95 Ala
Thr Ala Pro Ile Ile Ser Leu Phe Val Gly Phe Phe Trp Pro Lys
100 105 110 Lys Asp Lys Gly Pro
Gln Leu Ile Asp Ile Ile Asp Lys Glu Ile Lys 115
120 125 Lys Leu Leu Asp Lys Glu Leu Gly Glu
Gln Lys Arg Asn Asp Leu Val 130 135
140 Ser Ala Leu Asn Glu Met Gln Glu Gly Ala Asn Glu Leu
Ser Asp Ile145 150 155
160 Met Thr Asn Ala Leu Phe Glu Gly Thr Ile Gln Gly Asn Val Val Thr
165 170 175 Asn Asp Asn Pro
Gln Gly Lys Arg Arg Thr Pro Lys Ala Pro Thr Val 180
185 190 Ser Asp Tyr Glu Asn Val Tyr Ser Ala
Tyr Phe Val Glu His Val Asp 195 200
205 Phe Arg Asn Lys Ile Ser Thr Phe Leu Thr Gly Ser Tyr Asp
Leu Ile 210 215 220
Ala Leu Pro Leu Tyr Ala Leu Ala Lys Thr Met Glu Leu Ser Leu Tyr225
230 235 240 Gln Ser Phe Ile Asn
Phe Ala Asn Lys Trp Met Asp Phe Val Tyr Thr 245
250 255 Lys Ala Ile Asn Glu Ser Ala Thr Asp Asp
Met Lys Arg Asp Tyr Gln 260 265
270 Ala Arg Tyr Asn Thr Gln Lys Ser Asn Leu Ala Val Gln Lys Thr
Gln 275 280 285 Leu
Ile Asn Lys Ile Lys Asp Gly Thr Asp Ala Val Met Lys Val Phe 290
295 300 Lys Asp Thr Asn Asn Leu
Pro Ser Ile Gly Thr Asn Lys Leu Ala Val305 310
315 320 Asn Ala Arg Asn Lys Tyr Ile Arg Ala Leu Gln
Ile Asn Cys Leu Asp 325 330
335 Leu Val Ala Leu Trp Pro Gly Leu Tyr Pro Asp Glu Tyr Leu Leu Pro
340 345 350 Leu Gln Leu
Asp Lys Thr Arg Val Val Phe Ser Asp Thr Met Gly Pro 355
360 365 Asp Glu Thr His Asp Gly Gln Met
Lys Val Leu Asn Ile Leu Asp Ser 370 375
380 Thr Thr Ser Tyr Asn His Gln Asp Ile Gly Ile Ser Thr
Thr Gln Asp385 390 395
400 Val Asn Ser Leu Leu Phe Tyr Pro Arg Lys Glu Leu Leu Glu Leu Asp
405 410 415 Phe Ala Lys Tyr
Ile Ser Ser Ser Ser Arg Phe Trp Val Tyr Gly Phe 420
425 430 Gly Leu Lys Tyr Ser Asp Asp Asn Phe
Tyr Arg Tyr Gly Asp Asn Asp 435 440
445 Pro Ser Ser Asp Phe Lys Pro Ala Tyr Lys Trp Phe Thr Lys
Asn Ser 450 455 460
Gln Phe Glu Asn Leu Pro Thr Tyr Gly Asn Pro Thr Pro Ile Thr Asn465
470 475 480 Leu Asn Ala Lys Thr
Gln Val Thr Ser Tyr Leu Asp Ala Leu Ile Tyr 485
490 495 Tyr Ile Asp Gly Gly Thr Asn Leu Tyr Asn
Asn Ala Ile Leu His Asp 500 505
510 Thr Gly Gly Tyr Ile Pro Gly Tyr Pro Gly Val Glu Gly Tyr Gly
Met 515 520 525 Ser
Asn Asn Glu Pro Leu Ala Gly Gln Lys Leu Asn Ala Leu Tyr Pro 530
535 540 Ile Lys Val Glu Asn Val
Ser Gly Ser Gln Gly Lys Leu Gly Thr Ile545 550
555 560 Ala Ala Tyr Val Pro Leu Asn Leu Gln Pro Glu
Asn Ile Ile Gly Asp 565 570
575 Ala Asp Pro Asn Thr Gly Phe Pro Leu Asn Val Ile Lys Gly Phe Pro
580 585 590 Phe Glu Lys
Tyr Gly Pro Asp Tyr Glu Gly Arg Gly Ile Ser Val Val 595
600 605 Lys Glu Trp Ile Asn Gly Ala Asn
Ala Val Lys Leu Ser Pro Gly Gln 610 615
620 Ser Val Gly Val Gln Ile Lys Asn Ile Thr Lys Gln Asn
Tyr Gln Ile625 630 635
640 Arg Thr Arg Tyr Ala Ser Asn Asn Ser Asn Gln Val Tyr Phe Asn Val
645 650 655 Asp Pro Gly Gly
Ser Pro Leu Phe Ala Gln Ser Val Thr Phe Glu Ser 660
665 670 Thr Thr Asn Val Thr Ser Gly Gln Gln
Gly Glu Asn Gly Arg Tyr Thr 675 680
685 Leu Lys Thr Ile Phe Ser Gly Asn Asp Leu Leu Thr Val Glu
Ile Pro 690 695 700
Val Gly Asn Phe Tyr Val His Val Thr Asn Lys Gly Ser Ser Asp Ile705
710 715 720 Phe Leu Asp Arg Leu
Glu Phe Ser Thr Val Pro Ser Tyr Val Ile Tyr 725
730 735 Ser Gly Asp Tyr Asp Ala Thr Gly Thr Asp
Asp Val Leu Leu Ser Asp 740 745
750 Pro His Glu Tyr Phe Tyr Asp Val Ile Val Asn Gly Thr Ala Ser
His 755 760 765 Ser
Ser Ala Ala Thr Ser Met Asn Leu Leu Asn Lys Gly Thr Val Val 770
775 780 Arg Ser Ile Asp Ile Pro
Gly His Ser Thr Ser Tyr Ser Val Gln Tyr785 790
795 800 Ser Val Pro Glu Gly Phe Asp Glu Val Arg Ile
Leu Ser Ser Leu Pro 805 810
815 Asp Ile Ser Gly Thr Ile Arg Val Glu Ser Ser Lys Pro Pro Val Phe
820 825 830 Lys Asn Asp
Gly Asn Ser Gly Asp Gly Gly Asn Thr Glu Tyr Asn Phe 835
840 845 Asn Phe Asp Leu Ser Gly Leu Gln
Asp Thr Gly Leu Tyr Ser Gly Lys 850 855
860 Leu Lys Ser Gly Ile Arg Val Gln Gly Asn Tyr Thr Tyr
Thr Gly Ala865 870 875
880 Pro Ser Leu Asn Leu Val Val Tyr Arg Asn Asn Ser Val Val Ser Thr
885 890 895 Phe Pro Val Gly
Ser Pro Phe Asp Ile Thr Ile Thr Thr Glu Thr Asp 900
905 910 Lys Val Ile Leu Ser Leu Gln Pro Gln
His Gly Leu Ala Thr Val Thr 915 920
925 Gly Thr Gly Thr Ile Thr Ile Pro Asn Asp Lys Leu Ala Ile
Val Tyr 930 935 940
Asp Lys Leu Phe Lys Leu Pro His Asp Leu Glu Asn Ile Arg Ile Gln945
950 955 960 Val Asn Ala Leu Phe
Ile Ser Ser Thr Gln Asn Glu Leu Ala Lys Glu 965
970 975 Val Asn Asp His Asp Ile Glu Glu Val Ala
Leu Lys Val Asp Ala Leu 980 985
990 Ser Asp Glu Val Phe Gly Lys Glu Lys Lys Glu Leu Arg Lys Leu
Val 995 1000 1005 Asn
Gln Ala Lys Arg Leu Ser Lys Ala Arg Asn Leu Leu Val Gly Gly 1010
1015 1020 Asn Phe Asp Asn Trp Glu
Ala Trp Tyr Lys Gly Lys Glu Val Ala Arg1025 1030
1035 1040 Val Ser Asp His Glu Leu Leu Lys Ser Asp His
Val Leu Leu Pro Pro 1045 1050
1055 Pro Thr Met Tyr Pro Ser Tyr Ile Tyr Gln Lys Val Glu Glu Thr Lys
1060 1065 1070 Leu Lys Pro
Asn Thr Arg Tyr Met Ile Ser Gly Phe Ile Ala His Ala 1075
1080 1085 Glu Asp Leu Glu Ile Val Val Ser
Arg Tyr Gly Gln Glu Val Arg Lys 1090 1095
1100 Ile Val Gln Val Pro Tyr Gly Glu Ala Phe Pro Leu Thr
Ser Asn Gly1105 1110 1115
1120 Ser Ile Cys Cys Thr Pro Ser Phe Arg Arg Asp Gly Lys Leu Ser Asp
1125 1130 1135 Pro His Phe Phe
Ser Tyr Ser Ile Asp Val Gly Glu Leu Asp Met Thr 1140
1145 1150 Ala Gly Pro Gly Ile Glu Leu Gly Leu
Arg Ile Val Asp Arg Leu Gly 1155 1160
1165 Met Ala Arg Val Ser Asn Leu Glu Ile Arg Glu Asp Arg Ser
Leu Thr 1170 1175 1180
Ala Asn Glu Ile Arg Lys Val Gln Arg Met Ala Arg Asn Trp Arg Thr1185
1190 1195 1200 Glu Tyr Glu Lys Glu
Arg Ala Glu Val Thr Ala Leu Ile Glu Pro Val 1205
1210 1215 Leu Asn Gln Ile Asn Ala Leu Tyr Glu Asn
Gly Asp Trp Asn Gly Ser 1220 1225
1230 Ile Arg Ser Asp Ile Ser Tyr Tyr Asp Ile Glu Ser Ile Val Leu
Pro 1235 1240 1245 Thr
Leu Pro Arg Leu Arg His Trp Phe Val Pro Asp Met Leu Thr Glu 1250
1255 1260 His Gly Asn Ile Met Asn
Arg Phe Glu Glu Ala Leu Asn Arg Ala Tyr1265 1270
1275 1280 Thr Gln Leu Glu Gly Asn Thr Leu Leu His Asn
Gly His Phe Thr Thr 1285 1290
1295 Asp Ala Val Asn Trp Met Ile Gln Gly Asp Ala His Gln Val Ile Leu
1300 1305 1310 Glu Asp Gly
Arg Arg Val Leu Arg Leu Pro Asp Trp Ser Ser Ser Val 1315
1320 1325 Ser Gln Thr Ile Glu Ile Glu Lys
Phe Asp Pro Asp Lys Glu Tyr Asn 1330 1335
1340 Leu Val Phe His Ala Gln Gly Glu Gly Thr Val Thr Leu
Glu His Gly1345 1350 1355
1360 Glu Lys Thr Lys Tyr Ile Glu Thr His Thr His His Phe Ala Asn Phe
1365 1370 1375 Thr Thr Ser Gln
Ser Gln Gly Ile Thr Phe Glu Ser Asn Lys Val Thr 1380
1385 1390 Val Glu Ile Ser Ser Glu Asp Gly Glu
Leu Leu Val Asp His Ile Ala 1395 1400
1405 Leu Val Glu Val Pro Met Phe Asn Lys Asn Gln Met Val Asn
Glu Asn 1410 1415 1420
Arg Asp Val Asn Ile Asn Ser Asn Thr Asn Met Asn Asn Ser Asn Asn1425
1430 1435 1440 Gln35731PRTBacillus
thuringiensis 35Met Thr Thr Ile Asn Glu Leu Tyr Pro Ala Val Pro Tyr Asn
Val Leu1 5 10 15
Ala Tyr Ala Pro Pro Leu Asn Leu Ala Asp Ser Thr Pro Trp Gly Gln
20 25 30 Ile Val Val Ala Asp
Ala Ile Lys Glu Ala Trp Asp Asn Phe Gln Lys 35 40
45 Tyr Gly Val Leu Asp Leu Thr Ala Ile Asn
Gln Gly Phe Asp Asp Ala 50 55 60
Asn Thr Gly Ser Phe Ser Tyr Gln Ala Leu Ile Gln Thr Val Leu
Gly65 70 75 80 Ile
Ile Gly Thr Ile Gly Met Thr Val Pro Val Ala Ala Pro Phe Ala
85 90 95 Ala Thr Ala Pro Ile Ile
Ser Leu Phe Val Gly Phe Phe Trp Pro Lys 100
105 110 Lys Asp Lys Gly Pro Gln Leu Ile Asp Ile
Ile Asp Lys Glu Ile Lys 115 120
125 Lys Leu Leu Asp Lys Glu Leu Gly Glu Gln Lys Arg Asn Asp
Leu Val 130 135 140
Ser Ala Leu Asn Glu Met Gln Glu Gly Ala Asn Glu Leu Ser Asp Ile145
150 155 160 Met Thr Asn Ala Leu
Phe Glu Gly Thr Ile Gln Gly Asn Val Val Thr 165
170 175 Asn Asp Asn Pro Gln Gly Lys Arg Arg Thr
Pro Lys Ala Pro Thr Val 180 185
190 Ser Asp Tyr Glu Asn Val Tyr Ser Ala Tyr Phe Val Glu His Val
Asp 195 200 205 Phe
Arg Asn Lys Ile Ser Thr Phe Leu Thr Gly Ser Tyr Asp Leu Ile 210
215 220 Ala Leu Pro Leu Tyr Ala
Leu Ala Lys Thr Met Glu Leu Ser Leu Tyr225 230
235 240 Gln Ser Phe Ile Asn Phe Ala Asn Lys Trp Met
Asp Phe Val Tyr Thr 245 250
255 Lys Ala Ile Asn Glu Ser Ala Thr Asp Asp Met Lys Arg Asp Tyr Gln
260 265 270 Ala Arg Tyr
Asn Thr Gln Lys Ser Asn Leu Ala Val Gln Lys Thr Gln 275
280 285 Leu Ile Asn Lys Ile Lys Asp Gly
Thr Asp Ala Val Met Lys Val Phe 290 295
300 Lys Asp Thr Asn Asn Leu Pro Ser Ile Gly Thr Asn Lys
Leu Ala Val305 310 315
320 Asn Ala Arg Asn Lys Tyr Ile Arg Ala Leu Gln Ile Asn Cys Leu Asp
325 330 335 Leu Val Ala Leu
Trp Pro Gly Leu Tyr Pro Asp Glu Tyr Leu Leu Pro 340
345 350 Leu Gln Leu Asp Lys Thr Arg Val Val
Phe Ser Asp Thr Met Gly Pro 355 360
365 Asp Glu Thr His Asp Gly Gln Met Lys Val Leu Asn Ile Leu
Asp Ser 370 375 380
Thr Thr Ser Tyr Asn His Gln Asp Ile Gly Ile Ser Thr Thr Gln Asp385
390 395 400 Val Asn Ser Leu Leu
Phe Tyr Pro Arg Lys Glu Leu Leu Glu Leu Asp 405
410 415 Phe Ala Lys Tyr Ile Ser Ser Ser Ser Arg
Phe Trp Val Tyr Gly Phe 420 425
430 Gly Leu Lys Tyr Ser Asp Asp Asn Phe Tyr Arg Tyr Gly Asp Asn
Asp 435 440 445 Pro
Ser Ser Asp Phe Lys Pro Ala Tyr Lys Trp Phe Thr Lys Asn Ser 450
455 460 Gln Phe Glu Asn Leu Pro
Thr Tyr Gly Asn Pro Thr Pro Ile Thr Asn465 470
475 480 Leu Asn Ala Lys Thr Gln Val Thr Ser Tyr Leu
Asp Ala Leu Ile Tyr 485 490
495 Tyr Ile Asp Gly Gly Thr Asn Leu Tyr Asn Asn Ala Ile Leu His Asp
500 505 510 Thr Gly Gly
Tyr Ile Pro Gly Tyr Pro Gly Val Glu Gly Tyr Gly Met 515
520 525 Ser Asn Asn Glu Pro Leu Ala Gly
Gln Lys Leu Asn Ala Leu Tyr Pro 530 535
540 Ile Lys Val Glu Asn Val Ser Gly Ser Gln Gly Lys Leu
Gly Thr Ile545 550 555
560 Ala Ala Tyr Val Pro Leu Asn Leu Gln Pro Glu Asn Ile Ile Gly Asp
565 570 575 Ala Asp Pro Asn
Thr Gly Phe Pro Leu Asn Val Ile Lys Gly Phe Pro 580
585 590 Phe Glu Lys Tyr Gly Pro Asp Tyr Glu
Gly Arg Gly Ile Ser Val Val 595 600
605 Lys Glu Trp Ile Asn Gly Ala Asn Ala Val Lys Leu Ser Pro
Gly Gln 610 615 620
Ser Val Gly Val Gln Ile Lys Asn Ile Thr Lys Gln Asn Tyr Gln Ile625
630 635 640 Arg Thr Arg Tyr Ala
Ser Asn Asn Ser Asn Gln Val Tyr Phe Asn Val 645
650 655 Asp Pro Gly Gly Ser Pro Leu Phe Ala Gln
Ser Val Thr Phe Glu Ser 660 665
670 Thr Thr Asn Val Thr Ser Gly Gln Gln Gly Glu Asn Gly Arg Tyr
Thr 675 680 685 Leu
Lys Thr Ile Phe Ser Gly Asn Asp Leu Leu Thr Val Glu Ile Pro 690
695 700 Val Gly Asn Phe Tyr Val
His Val Thr Asn Lys Gly Ser Ser Asp Ile705 710
715 720 Phe Leu Asp Arg Leu Glu Phe Ser Thr Val Pro
725 730 36364PRTBacillus thuringiensis
36Met Leu Ile Lys Glu Met Gln Tyr Met His Ser Ile Lys Lys Tyr Lys1
5 10 15 Lys Val Leu Leu
Ile Ala Pro Leu Ala Cys Met Leu Thr Gly Ala Ile 20
25 30 Leu Pro Thr Ala Thr Thr Val His Ala
Gln Glu Val Glu Asn Lys Lys 35 40
45 Ala Val Ser Met Met Lys Pro Gly Gly Glu Phe Gly Ala Thr
Lys Tyr 50 55 60
Ser Lys Glu Asn Leu Val Lys Glu Ile Asn Leu Arg Leu Leu Thr Ala65
70 75 80 Leu Asp Arg Ser Thr
Ser Leu Arg Glu Lys Phe His Ile Lys Gly Asn 85
90 95 Glu Val Leu Asp Val Ser Gln Leu Asp Asp
Thr Ser Lys Gln Leu Met 100 105
110 Glu Lys Leu Gln Leu Thr Ala Glu Gly Ser Ile Asp Val Lys Pro
His 115 120 125 Val
Asp Ser Tyr Lys Asp Leu Gly Gln Thr Asn Ile Val Thr Tyr Asn 130
135 140 Asn Asp Asn Gly Val Val
Gly Gln Thr Tyr Asn Thr Pro Glu Thr Thr145 150
155 160 Val Lys Glu Ser Glu Thr His Thr Tyr Ser Asn
Thr Glu Gly Val Lys 165 170
175 Leu Gly Leu Glu Val Gly Thr Lys Ile Thr Val Gly Ile Pro Phe Ile
180 185 190 Gly Lys Asp
Glu Thr Glu Ile Lys Ala Thr Ser Glu Phe Ser Tyr Glu 195
200 205 His Asn Asp Ser Gln Thr Lys Thr
Lys Glu Thr Asp Val Thr Phe Lys 210 215
220 Ser Gln Pro Val Val Ala Ala Pro Gly Gly Thr Thr Thr
Tyr Tyr Gly225 230 235
240 Asp Ile Lys Thr Ala Thr Phe Ser Gly Ser Phe Gln Ser Asp Ala Tyr
245 250 255 Val Ala Gly Gly
Phe Glu Leu Lys Val Pro Ile Ala His Asp Met Ala 260
265 270 Ser Pro Lys Ile Asp Arg Tyr Glu Thr
Ala Thr Leu Thr Ala Ala Asp 275 280
285 Ile Tyr Glu Ile Phe Asn Ala Ser Asn Ala Ile Ala Ala Pro
Asn Tyr 290 295 300
Leu Lys Leu Asp Asn Ala Gly Lys Lys Val Leu Leu Thr Asp Lys Ala305
310 315 320 Thr Phe Asp Ile Asn
Gly Gln Gly Gly Phe Tyr Thr Thr Leu Gln Val 325
330 335 Lys Phe Val Pro Lys Asp Ser Asn Lys Lys
Pro Gln Met Met Ser Tyr 340 345
350 Lys Glu Tyr Val Gln Lys Met Asn Asn Asn Glu Leu 355
360 37394PRTBacillus thuringiensis 37Met
Tyr Ser Ile Lys Arg Tyr Lys Lys Val Ala Ile Val Ala Pro Leu1
5 10 15 Val Cys Leu Leu Gly Thr
Gly Leu Thr Phe Val Asn Lys Pro Ile Pro 20 25
30 Ala Ala Ala Ala Val Thr Thr Asn Tyr Ser Thr
Ala Asp Ser Ala Ser 35 40 45
Asn Phe Gln Pro Ile Ser Lys Tyr Thr Leu Ala Gly Asp Leu Tyr Glu
50 55 60 Arg Tyr Met
Arg Ala Leu Val Arg His Pro Glu Leu Leu Ser Ser Gly65 70
75 80 Gly Leu Lys Pro Val Thr Asn Gln
Thr Asp Leu Glu Gln Ile Asp Gly 85 90
95 Tyr Tyr Lys Val Met Ala Gln Phe Ile Arg Asp Asn Asn
Gln Asn Phe 100 105 110
Pro Ser Pro Phe Asn Arg Pro Ser Met Lys Leu Met Thr Gly Val Asn
115 120 125 Pro Phe Phe Asn
Trp Ala Pro Gln Tyr Thr Asn Leu Ser Thr Gln Asn 130
135 140 Val Ile Asn Leu Asp Asn Pro Lys
Val Asp Asp Tyr Lys Glu Asp Asn145 150
155 160 Ile Glu Leu Ala Thr Tyr Thr Asn Asn Thr Thr Ser
Glu Gln Thr Phe 165 170
175 Ser Thr Pro Ser Lys Ser Glu Lys Val Thr Asp Ser Phe Thr Tyr Ser
180 185 190 Asn Ser Glu
Gly Gly Lys Leu Gly Val Ser Ser Thr Thr Thr Ile Arg 195
200 205 Ala Gly Ile Pro Ile Ala Gln Ala
Gln Glu Thr Leu Thr Met Ser Phe 210 215
220 Glu Ala Thr Tyr Asn His Thr Ser Ser Asn Thr Ser Ser
Thr Glu Lys225 230 235
240 Thr Val Thr Tyr Pro Ser Gln Val Leu Lys Cys Leu Pro Gly Tyr Arg
245 250 255 Thr Ser Leu Ile
Val Lys Val Ser Gln Ala Asn Phe Ser Gly Thr Met 260
265 270 Asp Phe Asp Val Glu Pro Thr Val Ser
Ser Leu Ile Asp Gly Ile Glu 275 280
285 Lys Asn Trp Lys Asp Ile Lys Asp Asp Lys Thr Ile Lys Gly
Asp Lys 290 295 300
Ser Gly Asp Tyr Thr Val Pro Asn Arg Gln Glu Phe Leu Tyr Asn Val305
310 315 320 Tyr Lys Tyr Ser Asp
Leu Pro Ile Pro Ser Tyr Val Lys Leu Asp Asp 325
330 335 Lys Lys Lys Thr Val Ser Phe Gly Lys Val
Thr Thr Pro Tyr Thr Gly 340 345
350 Val Ala Gly His Leu Ser Glu Ala Asn Ala Thr Gln Val Lys Leu
Glu 355 360 365 Ser
Leu Asp Lys Ala Gln Lys Pro Ile Ile Met Pro Leu Lys Gln Tyr 370
375 380 Gln Gln Lys Ile Gln Asn
His Glu Ser Phe385 390 38360PRTBacillus
thuringiensis 38Met His Ser Ile Lys Lys Tyr Lys Lys Ile Leu Leu Val Ala
Pro Leu1 5 10 15
Ala Cys Met Leu Thr Gly Ala Ile Leu Pro Thr Ala Thr Thr Val His
20 25 30 Ala Gln Glu Ile Lys
Gly Pro Gly Val Met Lys Pro Asp Val Pro Trp 35 40
45 Asn Gln Glu His Tyr Thr Lys Glu Asn Leu
Ala Trp Arg Ala Ala Asp 50 55 60
Arg Leu Ser Tyr Ala Ala Asp Arg Ile Pro Ser Leu Arg Glu Lys
Phe65 70 75 80 Lys
Leu Lys Pro Asn Glu His Phe Tyr Cys Ser Asn Asp Thr Arg Tyr
85 90 95 Tyr Met Glu Glu Thr Leu
Leu Lys Asn Leu Gln Leu Ser Ala Glu Gly 100
105 110 Pro Ile Asn Val Thr Pro His Val Asp Ser
Tyr Thr Asp Leu Gly Gln 115 120
125 Thr Asn Leu Leu Thr Tyr Asn Asn Asp Asp Gly Ile Val Glu
Gln Lys 130 135 140
Ala Ser Thr Pro Glu Thr Thr Ile Lys Glu Ser Glu Thr Ser Ser Tyr145
150 155 160 Ser Asn Lys Glu Gly
Val Thr Leu Gly Ala Glu Val Glu Ser Lys Val 165
170 175 Thr Phe Asn Ile Pro Phe Ile Val Ala Gly
Glu Thr Lys Val Ile Ala 180 185
190 Lys Ser Glu Phe Ser Tyr Glu His Asp Asp Thr Gln Thr Lys Thr
His 195 200 205 Glu
Lys Glu Val Thr Phe Lys Ser Gln Glu Ile Val Ala Ala Pro Glu 210
215 220 Gly Thr Thr Thr Tyr Tyr
Gly Ser Ile Lys Thr Ala Asn Phe Ser Gly225 230
235 240 Ser Phe Gln Ser Asp Ala Val Val Gly Gly Gly
Val Thr Leu Thr Leu 245 250
255 Pro Ile Gly Val Met Asp Lys Asp Gly Gly Gln Lys Lys Thr His Thr
260 265 270 Glu Thr Ala
Thr Leu Thr Ala Glu Asp Met Tyr Glu Ile Phe Lys Ala 275
280 285 Pro Met Pro Trp Asp Met Asn Lys
Leu Pro Pro Tyr Leu Lys Leu Asp 290 295
300 Asp Ser Gly Lys Arg Val Leu Leu Ala Glu Lys Ala Thr
Phe Asp Ile305 310 315
320 Lys Gly Gln Gly Gly Phe Tyr Thr Glu Ile Gln Ala Lys Phe Val Pro
325 330 335 Lys Asp Lys Asn
Lys Lys Thr Gln Ile Met Pro Tyr Ala Glu Tyr Val 340
345 350 Gln Lys Val Lys Gln Asn Ala Leu
355 360 39676PRTBacillus thuringiensis 39Met Lys Ser
Met Asn Ser Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Leu1 5
10 15 Asp Ala Ser Gln Asn Asn Ser Thr
Met Ser Thr Arg Tyr Pro Arg Tyr 20 25
30 Pro Leu Ala Lys Asp Pro Gln Ala Ser Met Gln Thr Thr
Asn Tyr Lys 35 40 45
Asp Trp Leu Asn Leu Cys Asp Thr Pro Asn Met Glu Asn Pro Glu Phe 50
55 60 Gln Ser Val Gly Arg
Ser Ala Leu Ser Ile Leu Ile Asn Leu Ser Ser65 70
75 80 Arg Ile Leu Ser Leu Leu Gly Ile Pro Phe
Ala Ala Gln Ile Gly Gln 85 90
95 Leu Trp Ser Tyr Thr Leu Asn Leu Leu Trp Pro Val Ala Asn Asn
Ala 100 105 110 Thr
Gln Trp Glu Ile Phe Met Arg Thr Ile Glu Glu Leu Ile Asn Ala 115
120 125 Arg Ile Glu Thr Ser Val
Arg Asn Arg Ala Leu Ala Glu Leu Ala Gly 130 135
140 Leu Gly Asn Ile Leu Glu Asp Tyr Lys Val Val
Leu Gln Arg Trp Asn145 150 155
160 Leu Asn Pro Thr Asn Pro Thr Leu Gln Arg Asp Val Val Arg Gln Phe
165 170 175 Glu Ile Val
His Ala Phe Phe Arg Phe Gln Met Pro Val Phe Ala Val 180
185 190 Asp Gly Phe Glu Val Pro Leu Leu
Pro Val Tyr Ala Ser Ala Ala Asn 195 200
205 Leu His Leu Leu Leu Leu Arg Asp Val Val Ile Asn Gly
Ala Arg Trp 210 215 220
Gly Leu Glu Ser Asp Val Ile Asn Asp Tyr His Asp Leu Gln Leu Arg225
230 235 240 Leu Thr Ser Thr Tyr
Val Asp His Cys Val Thr Trp Tyr Asn Thr Gly 245
250 255 Leu Asn Arg Leu Ile Gly Thr Asn Ala Arg
Gln Trp Val Thr Tyr Asn 260 265
270 Gln Phe Arg Arg Glu Met Thr Ile Ser Val Leu Asp Ile Ile Ser
Leu 275 280 285 Phe
Ser Asn Tyr Asp Val Arg Arg Tyr Pro Thr Lys Thr Gln Ser Glu 290
295 300 Leu Thr Arg Met Ile Tyr
Thr Asp Pro Ile Gly Thr Glu Gly Asn Gln305 310
315 320 Phe Ile Pro Gly Trp Val Asp Asn Ala Pro Ser
Phe Ser Val Ile Glu 325 330
335 Asn Ser Val Val Arg Ser Pro Gly Ala Phe Thr Phe Leu Glu Arg Val
340 345 350 Gly Ile Phe
Thr Gly Phe Leu His Gly Trp Ser Ser Arg Ser Glu Phe 355
360 365 Trp Ser Ala His Arg Leu Phe Ser
Arg Pro Val Leu Gly Trp Ile Trp 370 375
380 Glu Ser Val Ile Phe Gly Asn Pro Gln Asn Asn Ile Gly
Tyr Gln Glu385 390 395
400 Val Asp Phe Thr Asn Phe Asp Val Phe Ser Ile Asn Ser Arg Ala Thr
405 410 415 Ser His Met Phe
Pro Asn Gly Ser Ala Arg Leu Phe Gly Val Pro Arg 420
425 430 Val Thr Phe Asp Leu Ser Asn Val Thr
Asn Asn Asn Leu Ala Gln Arg 435 440
445 Thr Tyr Asn Arg Pro Phe Thr Phe Gly Gly Gln Asp Ile Val
Ser Arg 450 455 460
Leu Pro Gly Glu Thr Thr Glu Ile Pro Asn Ser Ser Asn Phe Ser His465
470 475 480 Arg Leu Ala His Ile
Ser Ser Phe Pro Val Gly Asn Asn Gly Ser Val 485
490 495 Leu Ser Tyr Gly Trp Thr His Arg Asn Val
Asn Arg His Asn Arg Leu 500 505
510 Asn Pro Asn Ser Ile Thr Gln Ile Pro Ala Ile Lys Phe Ala Ser
Gly 515 520 525 Ser
Ala Arg Arg Gly Pro Gly His Thr Gly Gly Asp Leu Ala Ile Ala 530
535 540 Gln Gln His Ser Gly Tyr
Gln Leu Phe Met Gln Ser Pro Ser Ala Gln545 550
555 560 Arg Tyr Arg Leu Arg Leu Arg Tyr Ala Gly Ile
Ser Gly Gly Ser Ile 565 570
575 Ser Val Ser His Arg Asp Glu Asn Asn Gln Asn Ile Leu His Ser Ala
580 585 590 Thr Phe Asn
Val Arg Ala Thr Ser Gly Gln Leu Arg Tyr Ala Asp Phe 595
600 605 Ile Tyr Thr Asp Leu Glu Glu Asn
Thr Thr Leu Phe Glu Thr Arg Asn 610 615
620 Gly Val Asn Leu Tyr Arg Leu Met Ile Phe Val Ser Ser
Gly Ser Ile625 630 635
640 Leu Ile Asp Arg Ile Glu Tyr Ile Pro Glu Asn Thr Thr Thr Ile Glu
645 650 655 Tyr Glu Glu Glu
Arg Asn Leu Glu Lys Glu Lys Lys Ala Val Asp Asp 660
665 670 Leu Phe Thr Asn 675
40567PRTBacillus thuringiensis 40Met Asn Asn Met Tyr Thr Asn Asn Met Lys
Thr Thr Leu Lys Leu Glu1 5 10
15 Thr Thr Asp Tyr Glu Ile Asp Gln Ala Ala Ile Ser Ile Glu Cys
Met 20 25 30 Ser
Asp Glu Gln Asp Leu Gln Glu Lys Met Met Leu Trp Asp Glu Val 35
40 45 Lys Leu Ala Lys Gln Leu
Ser Gln Ser Arg Asn Leu Leu Tyr Asn Gly 50 55
60 Asp Phe Glu Asp Ser Ser Asn Gly Trp Lys Thr
Ser Asn Asn Ile Thr65 70 75
80 Ile Gln Leu Glu Asn Pro Ile Leu Lys Gly Lys Tyr Leu Asn Met Pro
85 90 95 Gly Ala Arg
Asp Ile Tyr Gly Thr Ile Phe Pro Thr Tyr Val Tyr Gln 100
105 110 Lys Ile Asp Glu Ser Lys Leu Lys
Pro Asn Thr Arg Tyr Arg Val Arg 115 120
125 Gly Phe Val Gly Ser Ser Lys Asp Leu Lys Leu Val Val
Thr Arg Tyr 130 135 140
Glu Lys Glu Ile Asp Ala Ser Met Asp Val Pro Asn Asp Leu Ser Tyr145
150 155 160 Met Gln Pro Ser Pro
Ser Cys Gly Asp Tyr Gly Cys Asp Ser Ser Ser 165
170 175 Gln Pro Met Met Asn Gln Gly Tyr Pro Thr
Pro Tyr Thr Asp Asp Tyr 180 185
190 Ala Ser Asp Met Tyr Ala Cys Ser Ser Asn Leu Gly Lys Lys His
Val 195 200 205 Lys
Cys His Asp Arg His Pro Phe Asp Phe His Ile Asp Thr Gly Glu 210
215 220 Leu Asp Thr Asn Thr Asn
Leu Gly Ile Cys Ile Leu Phe Lys Ile Ser225 230
235 240 Asn Pro Asp Gly Tyr Ala Thr Leu Gly Asn Leu
Glu Val Ile Glu Glu 245 250
255 Gly Pro Leu Thr Ser Glu Ala Leu Ala His Val Asn Gln Lys Glu Lys
260 265 270 Lys Trp Asn
Gln Gln Met Glu Lys Lys Arg Ser Glu Thr Gln Gln Ala 275
280 285 Tyr Asp Pro Ala Lys Gln Ala Val
Asp Ala Leu Phe Thr Asn Ser Gln 290 295
300 Gly Glu Glu Leu His Tyr His Ile Thr Leu Asp His Ile
Gln Asn Ala305 310 315
320 Asn Gln Leu Val Gln Ser Ile Pro Tyr Val His His Ala Trp Leu Pro
325 330 335 Asp Ala Pro Gly
Met Asn Tyr Asp Leu Tyr Asn Asn Leu Lys Val Arg 340
345 350 Ile Glu Gln Ala Arg Tyr Leu Tyr Asp
Ala Arg Asn Val Ile Thr Asn 355 360
365 Gly Asp Phe Ala Gln Gly Leu Thr Gly Trp His Ala Thr Gly
Lys Val 370 375 380
Asp Val Gln Gln Met Asp Gly Ala Ser Val Leu Val Leu Ser Asn Trp385
390 395 400 Ser Ala Gly Val Ser
Gln Asn Leu His Ala Gln Asp His His Gly Tyr 405
410 415 Met Leu Arg Val Ile Ala Lys Lys Glu Gly
Pro Gly Lys Gly Tyr Val 420 425
430 Thr Met Met Asp Cys Asn Gly His Gln Glu Thr Leu Lys Phe Thr
Ser 435 440 445 Cys
Glu Glu Gly Tyr Met Thr Lys Thr Val Glu Val Phe Pro Glu Ser 450
455 460 Asp Arg Val Arg Ile Glu
Ile Gly Glu Thr Glu Gly Thr Phe Tyr Ile465 470
475 480 Asp Ser Ile Glu Leu Leu Cys Met Gln Gly Tyr
Ala Ser Asn Asn Thr 485 490
495 Pro His Thr Gly Asn Met Tyr Glu Gln Ser Tyr Asn Gly Ile Tyr Asn
500 505 510 Gln Asn Thr
Ser Asp Leu Tyr His Gln Gly Tyr Thr Asn Asn Tyr Asn 515
520 525 Gln Glu Ser Ser Ser Met Tyr Asn
Gln Asn Tyr Thr Asn Asn Asp Asp 530 535
540 Gln His Ser Gly Cys Thr Cys Asn Gln Gly His Asn Ser
Gly Cys Thr545 550 555
560 Cys Asn Gln Gly Tyr Asn Arg 565
41640PRTBacillus thuringiensis 41Met Glu Glu Leu Glu Leu Lys Arg Thr Asn
Thr Leu Ser Ser Glu Asp1 5 10
15 Val Asn Ile Leu Gln Ile Glu Asn Leu Val Lys Glu Tyr Val Lys
Gln 20 25 30 Thr
Tyr Gly Asn Ser Ala Glu Ile Lys Lys Leu Ser Leu Asp Gly Leu 35
40 45 Asp Val Leu Tyr Asn Leu
Asp Ile Pro Ser Ile Leu Lys Gly Thr Ser 50 55
60 Ser Ser Ser Ala Ile Lys Val Gly Thr Asp Asn
Leu Asn Asn Pro Thr65 70 75
80 Asp Thr Ala Lys Thr Ile Lys Leu Pro Val Lys Asn Val Arg Lys Lys
85 90 95 Glu Phe Lys
Val Lys Pro Ile Gln Ala Leu Asn Phe Glu Asn Gly Ala 100
105 110 Thr Ile Thr Lys Lys Ser Ile Thr
Ser Ile Pro Ser Ile Asn Ala Thr 115 120
125 Phe Ile Ala Leu Ala Glu Gln Asn Phe Gln Asn Ala His
Phe His Ile 130 135 140
Val Asn Asp Ser Gln Ser Tyr Glu Asn Glu Ile Pro Ile Tyr Val Pro145
150 155 160 Pro His Ser Lys Val
Glu Ile Thr Tyr Tyr Val Lys Glu Ile Gln Phe 165
170 175 Asp Ala Ile Ile Gln Ser Thr Ala Thr Ile
Gly Gly Ser Ile Ser Phe 180 185
190 Glu Tyr Ile Val His Asp Asn Gly His Glu Gly Ile Asp Phe Leu
Thr 195 200 205 Ile
Phe Glu Leu Val Asn Ser Leu Asn Leu Asn Asp Phe Glu Ile Gln 210
215 220 Glu Ala Ser Asp Val His
Gly Lys Val Val Tyr Lys Gly Lys Ser Gln225 230
235 240 Phe Gln Gly Thr Val Gly Leu Asn Leu Phe Met
Gln Ile Lys Gly Thr 245 250
255 Pro Leu Asp Glu Ser Lys Asn Asn Tyr Glu Phe Thr Lys Val Leu Ser
260 265 270 Glu Asp Val
Glu Met Ser Leu Ser Pro Ser Glu Gly Glu Tyr Tyr Ile 275
280 285 Asp Phe Gly Ser Ser Pro Lys Leu
Thr Asn Lys Glu Glu Val Ile Val 290 295
300 Lys Phe Thr Arg Asp Tyr Leu Leu Ser Asn Asp Arg Lys
Asn Ala Tyr305 310 315
320 Val Gln Gln Leu Pro Arg Leu Glu Tyr Gly Glu Glu Val Thr Thr Leu
325 330 335 Lys Ser Ile Asp
Thr Ala His Glu Arg Lys Glu Ile Ile Ala Ser Thr 340
345 350 Ile Asn Thr Phe Gln Asn Pro Ser Asp
Thr Glu Ile Thr Arg Asn Thr 355 360
365 Ile Lys Glu Thr Phe Ser Thr Thr Asp Thr Ile Thr Thr Thr
Ala Thr 370 375 380
Thr Asp Lys Phe Leu Glu Leu Gly Gly Ser Ile Glu Thr Ser Ala Lys385
390 395 400 Gly Lys Val Pro Leu
Val Ala Glu Ala Ser Ile Lys Val Thr Gln Ser 405
410 415 Ile Lys Gly Gly Trp Lys Trp Val Ser Thr
Lys Thr Asn Thr Arg Thr 420 425
430 Asn Val His Thr Ile Glu Ile Pro Ser Gln Ser Ile Lys Ile Pro
Pro 435 440 445 His
Lys Met Trp Lys Tyr Gln Tyr Ile Leu Thr Lys Phe Glu Ser Ser 450
455 460 Gly Tyr Leu Ser Ser Ala
Trp Glu Ile Asn Thr Lys Glu Ser Met Ser465 470
475 480 Ala Pro Glu Val His Ile Gly Tyr Tyr Asn Lys
Asp Leu Gln Asn Pro 485 490
495 Arg Asn Ile Thr Gly Leu Ser Ala Asn Val Glu Ser Gly Asn Val Val
500 505 510 Gly Arg Val
Phe Glu Phe Asn Lys Phe Gln Pro Gly Gly Leu His Tyr 515
520 525 Lys Ile Leu Asn Ser Glu Asn Ile
Leu Asn Ala Thr Pro Tyr Gln Phe 530 535
540 Phe Lys Glu Leu Ala Lys Arg Val Asn Gln Tyr Pro Leu
Ile Gln Asn545 550 555
560 Asn Pro Arg Tyr Arg Arg Leu Gly Ile Leu Leu Gly Phe Gly Lys Asp
565 570 575 Ile Ser Gln Ile
Thr Trp Glu Pro Gln Ile His Tyr Asn Glu His Val 580
585 590 Leu Phe Asp Ala Glu Glu Leu Leu Asn
Val Leu Arg Phe Asp Asp Ile 595 600
605 Ala Asn Lys Val Tyr Ala Leu Asp Gly Gly Thr Pro Phe Thr
Val Ala 610 615 620
Val Gly His Glu Leu Leu Pro Lys Glu Ser Ile Glu Pro Leu Asn Asn625
630 635 640 42966PRTBacillus
thuringiensis 42Met Met Asn Met Ser Asn Thr Leu Ala Pro Tyr Asn Val Leu
Arg Ser1 5 10 15
Met Asp Met Pro Asn Ile Ser Gly Thr Lys Trp Asp Lys Gly Met Phe
20 25 30 Ile Asn Ala Leu Asp
Asn Thr Ser Phe Leu Leu Glu Leu Ile Glu Lys 35 40
45 Gly Ile Asn Asp Asp Asp Asp Val Leu Gly
Leu Leu Ser Phe Ile Gly 50 55 60
Leu Thr Ala Leu Glu Ala Ile Pro Ile Val Gly Gly Val Met Ser
Lys65 70 75 80 Leu
Val Ser Met Ile Phe Phe Pro Thr Lys Ser Ser Ile Asn Phe Gln
85 90 95 Lys Ile Trp Glu Gln Leu
Glu Lys Ala Ile Glu Gln Ile Val Asp Lys 100
105 110 Lys Ile Thr Glu Ala Met Met Ser Gln Leu
Met Gln Glu Ile Ala Gly 115 120
125 Leu Ala Asp Val Leu Glu Glu Tyr Arg Asn Ala Tyr Asp Leu
Tyr Asn 130 135 140
Gly Lys Lys Leu Phe Asn Ile Pro Asp Lys Met Thr Pro Gly Asp Tyr145
150 155 160 Leu Ile Asn Val Phe
Thr Thr Ala Asn Leu Gln Phe Ile Gln Arg Ile 165
170 175 Pro Thr Phe Gln Asn Ser Ile Tyr Asp Val
Val Phe Leu Pro Phe Phe 180 185
190 Val His Ala Ala Glu Met His Ile Leu Leu Ile Arg Asp Ala Ala
Ile 195 200 205 His
Gly Gln Glu Trp Gly Met Asp Glu Thr Val His Gln Lys Phe Lys 210
215 220 Arg Asp Leu Lys Thr Leu
Ile Asn Lys Tyr Ser Ser Tyr Leu Leu Ala225 230
235 240 Thr Tyr Lys Lys Gly Leu Lys Glu Ala Ser Glu
Lys Lys Leu Glu Asn 245 250
255 Asn Asp Phe Pro Thr Ser Asn Asn Gln His His Tyr Ile Asn Thr Val
260 265 270 Arg Trp Asn
Val Ile Asn Gln Tyr Lys Arg Gly Met Ala Leu Thr Val 275
280 285 Phe Asp Phe Ala Tyr Lys Trp Lys
Tyr Tyr Gln Glu Val Tyr Gln Asn 290 295
300 Asn Ile Thr Leu Asn Pro Ala Arg Thr Ile Tyr Ser Asp
Ile Ala Gly305 310 315
320 Ser Val Tyr Pro Tyr Glu Lys Thr Thr Asn Glu Ile Asp Asn Ile Ile
325 330 335 Lys Glu Gln Asn
Leu Lys Tyr Arg Gly Leu Leu Lys Glu Leu Leu Ile 340
345 350 Asn His Gly Asp Arg Ile Asp Ser Ile
Gln Ser Lys Tyr Ile Arg Asn 355 360
365 Asn Glu Ile Ile Asp Ser Asn Arg Thr Gly Gly Ala Gly Gly
Arg Ala 370 375 380
Thr Phe Phe Asp Leu Lys Ser Pro Ile Asn Asn Pro Phe Ile Gln Val385
390 395 400 Asn Met Trp Ser Glu
Leu Val Pro Phe Ser Leu Gly Phe Lys Tyr Tyr 405
410 415 Asn Gly Glu Glu Ser Lys Leu Ile Trp Gly
Gly Gly Thr Pro Gly Lys 420 425
430 His Lys Phe Gly Ser Tyr His Tyr Val Gly Asn Lys Val Ser Ser
Ile 435 440 445 Ile
Gly Phe Gly Lys Asn Gly Thr Gly Gly Phe Asn Ser Leu Asp Ala 450
455 460 Met Val Val Gly Phe Lys
Arg Asp Asp Tyr Ile Pro Glu Asn Arg Phe465 470
475 480 Val Gly Val Asn Lys Asn Gly Glu Pro Val Thr
Lys Val Ile Asp Ala 485 490
495 Glu Asn Phe Tyr Gln Glu Lys Phe Gln Ser Asn Ile Lys Met Ile Asp
500 505 510 Glu Pro Met
Phe Gly Glu Ala Val Leu Gln Phe Glu Asn Tyr Ser Asn 515
520 525 Asn Leu Asn Lys Asp Ser Tyr Val
Thr Tyr Gln Ile Asp Ala Lys Ile 530 535
540 Glu Gly Thr Tyr Glu Leu His Val Ile Ile Gly Ala Lys
Lys Gln Lys545 550 555
560 Asp Lys Ile Ala Phe Lys Met Ala Leu Asn Glu Lys Gln Pro Glu Lys
565 570 575 Phe Ile Thr Glu
Pro Phe Asn Ala Gly Asp Ile Trp Glu Gly Ile Ser 580
585 590 Leu Ser Glu Gly Leu Val Tyr Lys Arg
Ile Leu Leu Gly Asn Phe Gln 595 600
605 Leu Lys Lys Gly Met Asn Arg Ile Thr Ile His Asn Gly Val
Leu Gln 610 615 620
Thr Ser Ala Asn Ile Lys Thr Trp Asn Leu Ala Lys Leu Glu Leu Thr625
630 635 640 Leu Thr Ser Asp Ser
Leu Lys Asp Pro Asp Ile Thr Thr Leu Tyr Asp 645
650 655 Lys Asp Asn Tyr Ser Gly Thr Lys Lys Phe
Ile Phe Glu Asn Thr Ser 660 665
670 Arg Leu Lys Asp Phe Asn Asp Lys Thr Ser Ser Ile Lys Val Glu
Ser 675 680 685 His
Leu Ala Gly Ile Arg Ile Tyr Gln Asp Tyr Asn Tyr Lys Gly Lys 690
695 700 Ser Met Asp Leu Val Gly
Gly Glu Lys Ile Ser Leu Lys Asn His Ser705 710
715 720 Phe Asn Asn Arg Ala Ser Ser Val Lys Phe Ala
Asn Ile Val Leu Tyr 725 730
735 Asn Gln Asp Asn Tyr Gln Gly Ser Arg Lys Leu Val Phe Glu Asp Ile
740 745 750 Pro Asp Leu
Gly Lys Gln Ser Phe Asn Asp Lys Thr Ser Ser Ile Val 755
760 765 Val Ser Ser Asn Val Ser Gly Ala
Arg Leu Tyr Glu His Ala Tyr Tyr 770 775
780 Lys Gly Lys Tyr Val Asp Val Val Gly Gly Gln Lys Leu
Asn Leu Lys785 790 795
800 Asn His Val Leu Asn Lys Lys Ile Ser Ser Ile Lys Phe Phe Lys Glu
805 810 815 Gly Glu Val Leu
Asn Gly Val Tyr Gln Ile Ile Thr Ala Ile Asn Asn 820
825 830 Thr Ser Val Ile Asp Lys His Leu Glu
Asn Ser Asn Val His Leu Trp 835 840
845 Glu Asn Ala Glu Asn Lys Asn Gln Lys Trp Arg Ile Glu Tyr
Asp Val 850 855 860
Ala Lys Lys Ala Tyr Gln Ile Lys Asn Met Leu Asp Glu Lys Leu Val865
870 875 880 Leu Ser Thr His Glu
Leu Phe Pro Ile Phe Ser Ala Leu Tyr Cys Leu 885
890 895 Pro Asn Lys Gly Tyr Val Ser Gln Tyr Trp
Ile Phe Glu Tyr Val Gly 900 905
910 Asn Gly Tyr Tyr Ile Ile Lys Asn Lys Ala Tyr Pro Asp Trp Val
Leu 915 920 925 Asp
Val Asp Gly Leu Asn Ser Asp Asn Gly Thr Leu Ile Lys Leu His 930
935 940 Ser Gln His Asp Leu Thr
Asp Pro Leu Ile Asn Ala Gln Lys Phe Lys945 950
955 960 Leu Lys Asp Ile Asn Asn 965
43769PRTBacillus thuringiensis 43Met Met Asn Met Ser Asn Thr Leu Ala
Pro Tyr Asn Val Leu Arg Ser1 5 10
15 Met Asp Met Pro Asn Ile Ser Gly Thr Lys Trp Asp Lys Gly
Met Phe 20 25 30
Ile Asn Ala Leu Asp Asn Thr Ser Phe Leu Leu Glu Leu Ile Glu Lys 35
40 45 Gly Ile Asn Asp Asp
Asp Asp Val Leu Gly Leu Leu Ser Phe Ile Gly 50 55
60 Leu Thr Ala Leu Glu Ala Ile Pro Ile Val
Gly Gly Val Met Ser Lys65 70 75
80 Leu Val Ser Met Ile Phe Phe Pro Thr Lys Ser Ser Ile Asn Phe
Gln 85 90 95 Lys
Ile Trp Glu Gln Leu Glu Lys Ala Ile Glu Gln Ile Val Asp Lys
100 105 110 Lys Ile Thr Glu Ala
Met Met Ser Gln Leu Met Gln Glu Ile Ala Gly 115
120 125 Leu Ala Asp Val Leu Glu Glu Tyr Arg
Asn Ala Tyr Asp Leu Tyr Asn 130 135
140 Gly Lys Lys Leu Phe Asn Ile Pro Asp Lys Met Thr Pro
Gly Asp Tyr145 150 155
160 Leu Ile Asn Val Phe Thr Thr Ala Asn Leu Gln Phe Ile Gln Arg Ile
165 170 175 Pro Thr Phe Gln
Asn Ser Ile Tyr Asp Val Val Phe Leu Pro Phe Phe 180
185 190 Val His Ala Ala Glu Met His Ile Leu
Leu Ile Arg Asp Ala Ala Ile 195 200
205 His Gly Gln Glu Trp Gly Met Asp Glu Thr Val His Gln Lys
Phe Lys 210 215 220
Arg Asp Leu Lys Thr Leu Ile Asn Lys Tyr Ser Ser Tyr Leu Leu Ala225
230 235 240 Thr Tyr Lys Lys Gly
Leu Lys Glu Ala Ser Glu Lys Lys Leu Glu Asn 245
250 255 Asn Asp Phe Pro Thr Ser Asn Asn Gln His
His Tyr Ile Asn Thr Val 260 265
270 Arg Trp Asn Val Ile Asn Gln Tyr Lys Arg Gly Met Ala Leu Thr
Val 275 280 285 Phe
Asp Phe Ala Tyr Lys Trp Lys Tyr Tyr Gln Glu Val Tyr Gln Asn 290
295 300 Asn Ile Thr Leu Asn Pro
Ala Arg Thr Ile Tyr Ser Asp Ile Ala Gly305 310
315 320 Ser Val Tyr Pro Tyr Glu Lys Thr Thr Asn Glu
Ile Asp Asn Ile Ile 325 330
335 Lys Glu Gln Asn Leu Lys Tyr Arg Gly Leu Leu Lys Glu Leu Leu Ile
340 345 350 Asn His Gly
Asp Arg Ile Asp Ser Ile Gln Ser Lys Tyr Ile Arg Asn 355
360 365 Asn Glu Ile Ile Asp Ser Asn Arg
Thr Gly Gly Ala Gly Gly Arg Ala 370 375
380 Thr Phe Phe Asp Leu Lys Ser Pro Ile Asn Asn Pro Phe
Ile Gln Val385 390 395
400 Asn Met Trp Ser Glu Leu Val Pro Phe Ser Leu Gly Phe Lys Tyr Tyr
405 410 415 Asn Gly Glu Glu
Ser Lys Leu Ile Trp Gly Gly Gly Thr Pro Gly Lys 420
425 430 His Lys Phe Gly Ser Tyr His Tyr Val
Gly Asn Lys Val Ser Ser Ile 435 440
445 Ile Gly Phe Gly Lys Asn Gly Thr Gly Gly Phe Asn Ser Leu
Asp Ala 450 455 460
Met Val Val Gly Phe Lys Arg Asp Asp Tyr Ile Pro Glu Asn Arg Phe465
470 475 480 Val Gly Val Asn Lys
Asn Gly Glu Pro Val Thr Lys Val Ile Asp Ala 485
490 495 Glu Asn Phe Tyr Gln Glu Lys Phe Gln Ser
Asn Ile Lys Met Ile Asp 500 505
510 Glu Pro Met Phe Gly Glu Ala Val Leu Gln Phe Glu Asn Tyr Ser
Asn 515 520 525 Asn
Leu Asn Lys Asp Ser Tyr Val Thr Tyr Gln Ile Asp Ala Lys Ile 530
535 540 Glu Gly Thr Tyr Glu Leu
His Val Ile Ile Gly Ala Lys Lys Gln Lys545 550
555 560 Asp Lys Ile Ala Phe Lys Met Ala Leu Asn Glu
Lys Gln Pro Glu Lys 565 570
575 Phe Ile Thr Glu Pro Phe Asn Ala Gly Asp Ile Trp Glu Gly Ile Ser
580 585 590 Leu Ser Glu
Gly Leu Val Tyr Lys Arg Ile Leu Leu Gly Asn Phe Gln 595
600 605 Leu Lys Lys Gly Met Asn Arg Ile
Thr Ile His Asn Gly Val Leu Gln 610 615
620 Thr Ser Ala Asn Ile Lys Thr Trp Asn Leu Ala Lys Leu
Glu Leu Thr625 630 635
640 Leu Thr Ser Asp Ser Leu Lys Asp Pro Asp Ile Thr Thr Leu Tyr Asp
645 650 655 Lys Asp Asn Tyr
Ser Gly Thr Lys Lys Phe Ile Phe Glu Asn Thr Ser 660
665 670 Arg Leu Lys Asp Phe Asn Asp Lys Thr
Ser Ser Ile Lys Val Glu Ser 675 680
685 His Leu Ala Gly Ile Arg Ile Tyr Gln Asp Tyr Asn Tyr Lys
Gly Lys 690 695 700
Ser Met Asp Leu Val Gly Gly Glu Lys Ile Ser Leu Lys Asn His Ser705
710 715 720 Phe Asn Asn Arg Ala
Ser Ser Val Lys Phe Ala Asn Ile Val Leu Tyr 725
730 735 Asn Gln Asp Asn Tyr Gln Gly Ser Arg Lys
Leu Val Phe Glu Asp Ile 740 745
750 Pro Asp Leu Gly Lys Gln Ser Phe Asn Asp Lys Thr Ser Ser Ile
Val 755 760 765
Val44575PRTBacillus thuringiensis 44Met Asn Gly Asn Gly Arg His Asp Gly
Trp Asn Gln Asn Gln His Ile1 5 10
15 Glu Asn Gly Gln Met Asn Pro Asn His Ser Gly Ser Cys Lys
Cys Gly 20 25 30
Cys Gln Gln Asn Asn Asn Gly Ser Tyr Pro Ser Asn Glu Tyr Asn Ser 35
40 45 Asn Asn Asn Gly Ser
Tyr Pro Ser Asn Glu Tyr Asn Ser Asn Asn Asn 50 55
60 Gly Ser Tyr Pro Ser Asn Glu Tyr Asn Ser
Asn Asn Asn Gly Ser Tyr65 70 75
80 Pro Ser Asn Glu Tyr Asn Ser Asn Asn Asn Gly Ser Tyr Pro Ser
Asn 85 90 95 Glu
Tyr Asn Ser Asn Asn Asn Gly Ser Tyr Pro Ser Asn Glu Tyr Asn
100 105 110 Ser Asn Asn Asn Gly
Ser Tyr Pro Ser Asn Glu Tyr Asn Ser Asn Asn 115
120 125 Asn Gly Ser Tyr Pro Ser Asn Glu Tyr
Asn Ser Asn Asn Asn Gly Ser 130 135
140 Tyr Ser Ser Asn Glu Tyr Asn Ser Asn Asn Asn Gly Ser
Tyr Pro Ser145 150 155
160 Asn Glu Tyr Asn Ser Asn Asn Asn Gly Ser Tyr Pro Ser Asn Glu Tyr
165 170 175 Asn Ser Asn Asn
Asn Gly Ser Tyr Pro Ser Asn Glu Tyr Val Gly Gly 180
185 190 Tyr Ser Ile Gln Asp Gly Leu Pro Gln
Glu Ser Lys Gln Phe Gln Lys 195 200
205 Ile Ser Asn Met Asn Thr Arg Asp Asn His Arg Val Leu Asp
Ala Gln 210 215 220
Asp Thr Tyr Phe Gly Gln Leu Ile Asp Asn Arg Val Gly Asp Thr Cys225
230 235 240 Lys Tyr Val Glu His
Lys Asn Ser Val Ile Tyr Glu Leu Ser Arg Gln 245
250 255 Pro Val Tyr Thr Pro Asp Ser Gln Tyr Phe
Ile Phe Tyr Gln Met Asp 260 265
270 Asn Gly Asn Phe Ile Ile Ala Asn Lys Glu Asn Ser Arg Val Leu
Glu 275 280 285 Val
Ile Phe Ser Ser Val Asn Gly Phe Val Thr Ile Ser Asn Glu Phe 290
295 300 Asn Ala Thr Ser Asp Gln
Arg Phe Lys Val Val Arg Ser Lys Asn Asp305 310
315 320 Thr Phe Arg Leu Val Thr Glu Gly Asn Lys Thr
Leu Asn Ile Cys Gly 325 330
335 His Ser Phe Gln Tyr Asn Thr Lys Ile Thr Ala Val Asn Ala Asp Ile
340 345 350 Asp Gly Asp
Asn Tyr Leu Phe Gln Lys Ser Met Asp Lys Asp Thr Arg 355
360 365 Asp Leu Tyr Phe Gly Thr Ile Ser
Asn Lys Asn Pro Glu Ile Leu Asn 370 375
380 Asp Pro Arg Asn Leu Lys Ser Leu Asp Asp Leu Gly Asp
Glu Pro Arg385 390 395
400 Ala Phe Lys Gly Ala Ala Leu Leu Pro Ala Leu Phe Val Asn Asp Pro
405 410 415 Arg Tyr Ser Val
His Arg Arg Val Ser Asn Ser Pro Tyr Tyr Tyr Leu 420
425 430 Glu Tyr Thr Gln Tyr Trp His Arg Ile
Trp Thr Asp Val Leu Pro Ile 435 440
445 Asp Gly Tyr Gly Ala Trp Ile Glu Met Ile Gly Val Thr Asn
Asp Thr 450 455 460
Gln Val Asn Met Lys Asn Ile Met Asn Ile Thr Ile Thr Gly Lys Asp465
470 475 480 Leu Gly Val Asp Leu
Gly Ile Asp Leu Gly Leu Arg Phe Gly Asp Lys 485
490 495 Ser Phe Leu Phe Glu Gln Lys Ile Leu Ser
Gly Leu Ser Ile Arg Lys 500 505
510 Thr Asp Tyr Pro Asn Leu Gly Ile Asp Glu Arg Ala Met Tyr Gln
Arg 515 520 525 Asn
Asn Ser Asn Leu Lys Thr Arg Phe Val Arg Tyr Val Lys Lys His 530
535 540 Glu Phe Val Leu Arg Asp
Leu Asn Gly Ser Lys Val Ala Glu Pro Trp545 550
555 560 Ile Ile Thr Glu Asp Arg Ser Ile Thr Lys Glu
Tyr Ser Ser Asn 565 570
575 45559PRTBacillus thuringiensis 45Met Lys Tyr Lys Asn Arg Thr Arg Ala
Lys Cys Lys Tyr Lys Gln Ala1 5 10
15 Leu Leu Val Thr Val Ala Thr Met Thr Leu Gly Val Ser Thr
Leu Gly 20 25 30
Ser Asn Ala Ser Ala Phe Ala Asp Glu Lys Glu Lys Asn Val Ile Gln 35
40 45 Gln Lys Ser Pro Gly
Thr Tyr Tyr Glu Asp Ala Gln Lys Asn Leu Gly 50 55
60 Ser Leu Ala Arg Phe Asp Thr Trp Ala Gln
Asp Leu Gly Lys Thr Thr65 70 75
80 Gly Ala Gly Asn Tyr Lys Thr Thr Leu Gly Met Ala Glu Lys Leu
Leu 85 90 95 Pro
Thr Ile Tyr Asn Asp Leu Asn Ser Gly Asn Phe Asn Asn Thr Ala
100 105 110 Arg Ser Ile Thr Met
Leu Ser Thr Ala Leu Ile Pro Tyr Gly Gly Ala 115
120 125 Phe Ile Ser Pro Ile Ile Gly Ile Leu
Trp Pro Glu Asn Gly Pro Asn 130 135
140 Ile Lys Glu Met Leu Gln Glu Met Glu Asn Lys Leu Val
Gly Ile Met145 150 155
160 Asp Glu Lys Ile Glu Ala Lys Asp Leu Asp Asp Leu Glu Ala Ala Val
165 170 175 Lys Gly Leu Met
Val Ser Leu Lys Glu Phe Glu Asn Ser Leu Asn Gly 180
185 190 Asn Ile Gly Gly Glu Tyr Tyr Ser Ala
Leu Ala Asp Val Asp Ser Leu 195 200
205 Asn Arg Gly Arg Ile Thr Ala Ile Gln Lys Gly Phe Asn Asp
Leu Ile 210 215 220
Ser Ala Thr Ser Lys Pro Lys Phe Lys Ile Thr Glu Leu Pro Leu Tyr225
230 235 240 Thr Ile Ile Ala Thr
Ala His Leu Asn Phe Leu Asn Thr Val Glu Lys 245
250 255 Gln Gly Thr Ser Pro Lys Ile Asn Tyr Thr
Glu Ala Ala Leu Lys Asp 260 265
270 Leu Leu Gln Asn Met Lys Lys Asn His Lys Asp Tyr Ala Asp Tyr
Ile 275 280 285 Glu
Lys Thr Tyr Thr Glu Gly Glu Ala Arg Ile Asn Ser Lys Leu Glu 290
295 300 Asp Lys Gln Lys Ile Glu
Gln Asp Leu Ala Ala Val Asn Gln Lys Leu305 310
315 320 Ser Glu Met Pro Arg Lys Pro Lys Asn His Thr
His Glu Glu Glu Asn 325 330
335 Lys Phe Ile Ile Gln Lys Glu Lys Leu Tyr Ala Gln Gln Asp Ser Leu
340 345 350 Glu Lys Lys
Leu Ser Glu Tyr Asn Asp Leu Met Tyr Gln Lys Ser Asp 355
360 365 Phe Tyr Ser Lys Thr Lys Gly Ser
Glu Ala Phe Gln Ile Ala Ser Thr 370 375
380 Gly Lys Thr Ile Pro Thr Pro Ser Trp Val Lys Thr Glu
Gly Thr Trp385 390 395
400 Val Cys Glu Ala Gly Phe Trp Phe Tyr Ile Asp Ala Lys Gly Gln Lys
405 410 415 Lys Ser Asp Trp
Phe Asn Asp Lys Thr Pro Asp Gly Lys Asp Arg Trp 420
425 430 Tyr Tyr Leu Ser Thr Glu Thr Pro Arg
Leu Asp Asn Val Arg Gly Asn 435 440
445 Ala Tyr Val Gly Lys Gly Thr Met Leu Thr Gly Trp Phe His
Asp Thr 450 455 460
Arg Lys Asp Lys Gln Ile Ile Gly Val Asn Thr Lys Thr Thr Tyr Glu465
470 475 480 Tyr Trp Tyr Tyr Leu
Ser Pro Glu Lys Asn Leu Lys Asn Ser Ala Gly 485
490 495 Glu Leu Phe Lys Gln Gly Gln Met Met Thr
Lys Trp Val Glu Ile Lys 500 505
510 Asp Thr Lys Thr Gly Glu Pro His Trp Tyr Tyr Phe Asn Pro Asp
Asp 515 520 525 Gly
Ser Met Thr His Asp Lys Lys Ala Val Gln Ile Gly Asp Lys Lys 530
535 540 Tyr Asp Phe Gly Ser Asn
Gly Val Cys Thr Thr Pro Asn Gly Tyr545 550
555 461293PRTBrevibacillus laterosporus 46Met Asn Gln
Asn Gln Asn Lys Asn Glu Met Gln Ile Ile Glu Pro Ser1 5
10 15 Ser Asp Ser Phe Leu Tyr Ser His
Asn Asn Tyr Pro Tyr Ala Thr Asp 20 25
30 Pro Asn Thr Val Leu Glu Gly Arg Asn Tyr Lys Glu Trp
Leu Asn Lys 35 40 45
Cys Thr Asp Asn Tyr Thr Asp Ala Leu Gln Ser Pro Glu Ala Thr Ala 50
55 60 Ile Ser Lys Gly Ala
Val Ser Ala Ala Ile Ser Ile Ser Thr Lys Val65 70
75 80 Leu Gly Leu Leu Gly Val Pro Phe Ala Ala
Gln Ile Gly Gln Leu Trp 85 90
95 Thr Phe Ile Leu Asn Ala Leu Trp Pro Ser Asp Asn Thr Gln Trp
Glu 100 105 110 Glu
Phe Met Arg His Val Glu Glu Leu Ile Asn Gln Arg Ile Ala Asp 115
120 125 Tyr Ala Arg Asn Lys Ala
Leu Ala Glu Leu Thr Gly Leu Gly Asn Asn 130 135
140 Leu Asp Leu Tyr Ile Glu Ala Leu Asp Asp Trp
Lys Arg Asn Pro Thr145 150 155
160 Ser Gln Glu Ala Lys Thr Arg Val Ile Asp Arg Phe Arg Ile Val Asp
165 170 175 Gly Leu Phe
Glu Ala Tyr Ile Pro Ser Phe Ala Val Ser Gly Tyr Gln 180
185 190 Val Gln Leu Leu Thr Val Tyr Ala
Ala Ala Ala Asn Leu His Leu Leu 195 200
205 Leu Leu Arg Asp Ser Thr Ile Tyr Gly Ile Asp Trp Gly
Leu Ser Gln 210 215 220
Thr Asn Val Asn Asp Asn Tyr Asn Arg Gln Ile Arg Leu Thr Ala Thr225
230 235 240 Tyr Ala Asn His Cys
Thr Thr Trp Tyr Gln Thr Gly Leu Glu Arg Leu 245
250 255 Arg Gly Ser Asn Ala Ser Ser Trp Val Thr
Tyr Asn Arg Phe Arg Arg 260 265
270 Glu Met Thr Leu Thr Val Leu Asp Ile Cys Ser Leu Phe Ser Asn
Tyr 275 280 285 Asp
Tyr Arg Ser Tyr Pro Ala Glu Val Arg Gly Glu Ile Thr Arg Glu 290
295 300 Ile Tyr Thr Asp Pro Val
Gly Val Gly Trp Val Asp Ser Ala Pro Ser305 310
315 320 Phe Gly Glu Ile Glu Asn Leu Ala Ile Arg Ala
Pro Arg Thr Val Thr 325 330
335 Trp Leu Asn Ser Thr Arg Ile Phe Thr Gly Arg Leu Gln Gly Trp Ser
340 345 350 Gly Thr Asn
Asn Tyr Trp Ala Ala His Met Gln Asn Phe Ser Glu Thr 355
360 365 Asn Ser Gly Asn Ile Gln Phe Glu
Gly Pro Leu Tyr Gly Ser Thr Val 370 375
380 Gly Thr Ile His Arg Thr Asp Asp Tyr Asp Met Gly Asn
Arg Asp Ile385 390 395
400 Tyr Thr Ile Thr Ser Gln Ala Val Leu Gly Leu Trp Ala Thr Gly Gln
405 410 415 Arg Val Leu Gly
Val Ala Ser Ala Arg Phe Thr Leu Arg Asn Leu Phe 420
425 430 Asn Asn Leu Thr Gln Val Leu Val Tyr
Glu Asn Pro Ile Ser Ser Thr 435 440
445 Phe Gly Ser Ser Thr Leu Thr His Glu Leu Ser Gly Glu Asn
Ser Asp 450 455 460
Arg Pro Thr Ser Ser Asp Tyr Ser His Arg Leu Thr Ser Ile Thr Gly465
470 475 480 Phe Arg Ala Gly Ala
Asn Gly Thr Val Pro Val Phe Gly Trp Thr Ser 485
490 495 Ala Thr Val Asp Arg Asn Asn Ile Ile Glu
Arg Asn Lys Ile Thr Gln 500 505
510 Phe Pro Gly Val Lys Ser His Thr Leu Asn Asn Cys Gln Val Val
Arg 515 520 525 Gly
Thr Gly Phe Thr Gly Gly Asp Trp Leu Arg Pro Asn Asn Asn Gly 530
535 540 Thr Phe Arg Leu Thr Ile
Thr Ser Phe Ser Ser Gln Ser Tyr Arg Ile545 550
555 560 Arg Leu Arg Tyr Ala Thr Ser Val Gly Asn Thr
Ser Leu Val Ile Ser 565 570
575 Ser Ser Asp Ala Gly Ile Ser Ser Thr Thr Ile Pro Leu Thr Ser Thr
580 585 590 Ile Thr Ser
Leu Pro Gln Thr Val Pro Tyr Gln Ala Phe Arg Val Val 595
600 605 Asp Leu Pro Ile Thr Phe Thr Thr
Pro Thr Thr Gln Arg Asn Tyr Thr 610 615
620 Phe Asp Phe Arg Leu Gln Asn Pro Ser Asn Ala Asn Val
Phe Ile Asp625 630 635
640 Arg Phe Glu Phe Val Pro Ile Gly Gly Ser Leu Ser Glu Tyr Glu Thr
645 650 655 Lys His Gln Leu
Glu Lys Ala Arg Lys Ala Val Asn Asp Leu Phe Thr 660
665 670 Asn Glu Ser Lys Asn Val Leu Lys Lys
Asp Thr Thr Asp Tyr Asp Ile 675 680
685 Asp Gln Ala Ala Asn Leu Val Glu Cys Val Ser Asp Glu Cys
Ala Asn 690 695 700
Ala Lys Met Ile Leu Leu Asp Glu Val Lys Tyr Ala Lys Gln Leu Ser705
710 715 720 Glu Ala Arg Asn Leu
Leu Leu Asn Gly Asn Phe Glu Tyr Gln Asp Arg 725
730 735 Asp Gly Glu Asn Pro Trp Lys Thr Ser Pro
Asn Val Thr Ile Gln Glu 740 745
750 Asn Asn Pro Ile Phe Lys Gly Arg Tyr Leu Ser Met Ser Gly Ala
Asn 755 760 765 Asn
Ile Glu Ala Thr Asn Glu Ile Phe Pro Thr Tyr Val Tyr Gln Lys 770
775 780 Ile Asp Glu Ser Lys Leu
Lys Pro Tyr Thr Arg Tyr Lys Val Arg Gly785 790
795 800 Phe Val Gly Asn Ser Lys Asp Leu Glu Leu Leu
Val Thr Arg Tyr Asp 805 810
815 Glu Glu Val Asp Ala Ile Leu Asn Val Pro Asn Asp Ile Pro His Ala
820 825 830 Pro Pro Pro
Phe Cys Gly Glu Phe Asp Arg Cys Lys Pro His Ser Tyr 835
840 845 Pro Pro Ile Asn Pro Glu Cys His
His Asp Val Ile Asn Asn Ile Glu 850 855
860 Ile Ser Ser Pro Cys Gln His Asn Lys Met Val Asp Asn
Ala Asp Ile865 870 875
880 Ser Tyr Arg His Ser Arg Leu Ser Lys Lys His Gly Ile Cys His Glu
885 890 895 Ser His His Phe
Glu Phe His Ile Asp Thr Gly Lys Ile Asp Leu Val 900
905 910 Glu Asn Leu Gly Ile Trp Val Val Phe
Lys Ile Cys Ser Thr Asp Gly 915 920
925 Tyr Ala Thr Leu Asp Asn Leu Glu Val Ile Glu Glu Gly Pro
Leu Gly 930 935 940
Ala Glu Ser Leu Glu Arg Val Lys Arg Arg Glu Lys Lys Trp Lys His945
950 955 960 His Met Glu His Lys
Cys Ser Glu Thr Lys His Ala Tyr His Ala Ala 965
970 975 Lys Gln Ala Val Val Ala Leu Phe Thr Asn
Ser Lys Tyr Asp Arg Leu 980 985
990 Lys Phe Glu Thr Thr Ile Ser Asn Ile Leu Phe Ala Asp Tyr Leu
Val 995 1000 1005 Gln
Ser Ile Pro Tyr Val Tyr Asn Lys Trp Leu Pro Gly Val Pro Gly 1010
1015 1020 Met Asn Tyr Asp Ile Tyr
Thr Glu Leu Lys Asn Leu Phe Thr Gly Ala1025 1030
1035 1040 Phe Asn Leu Tyr Asp Gln Arg Asn Ile Ile Lys
Asn Gly Asp Phe Asn 1045 1050
1055 Arg Gly Leu Met His Trp His Ala Thr Pro His Ala Arg Val Glu Gln
1060 1065 1070 Ile Ile Asp
Asn Arg Ser Val Leu Val Leu Pro Asn Tyr Ala Ala Asn 1075
1080 1085 Val Ser Gln Glu Val Cys Leu Glu
His Asn Arg Gly Tyr Val Leu Arg 1090 1095
1100 Val Thr Ala Lys Lys Glu Gly Pro Gly Ile Gly Tyr Val
Thr Phe Ser1105 1110 1115
1120 Asp Cys Ala Asn His Ile Glu Lys Leu Thr Phe Thr Ser Cys Asp Tyr
1125 1130 1135 Gly Thr Asn Val
Val Pro Tyr Glu Gln Ser Asn Tyr Pro Thr Asp Gly 1140
1145 1150 Val Pro Tyr Gly Gln His Gly Cys Asn
Ile Asp Gly Val Pro Tyr Glu 1155 1160
1165 Gln Ser Gly Tyr Arg Thr Asp Gly Val Pro Tyr Glu Gln Ser
Gly Tyr 1170 1175 1180
Arg Thr Asp Gly Val Pro Tyr Glu Gln Ser Gly His Arg Thr Asp Gly1185
1190 1195 1200 Val Pro Tyr Glu Gln
Ser Gly Tyr Arg Thr Asp Gly Val Pro Cys Glu 1205
1210 1215 Gln His Gly Cys His Thr Asp Gly Leu Pro
His Ile Gln His Gly Cys 1220 1225
1230 Arg Thr Asp Gly Leu Pro His Ile Gln His Gly Cys Arg Thr Asp
Arg 1235 1240 1245 Ser
Arg Asp Glu Leu Leu Gly Tyr Val Thr Lys Thr Ile Asp Val Phe 1250
1255 1260 Pro Asn Thr Asp Lys Val
Arg Ile Asp Ile Gly Glu Thr Glu Gly Thr1265 1270
1275 1280 Phe Lys Val Glu Ser Val Glu Leu Ile Cys Met
Glu Glu 1285 1290
47647PRTBrevibacillus laterosporus 47Met Asn Gln Asn Gln Asn Lys Asn Glu
Met Gln Ile Ile Glu Pro Ser1 5 10
15 Ser Asp Ser Phe Leu Tyr Ser His Asn Asn Tyr Pro Tyr Ala
Thr Asp 20 25 30
Pro Asn Thr Val Leu Glu Gly Arg Asn Tyr Lys Glu Trp Leu Asn Lys 35
40 45 Cys Thr Asp Asn Tyr
Thr Asp Ala Leu Gln Ser Pro Glu Ala Thr Ala 50 55
60 Ile Ser Lys Gly Ala Val Ser Ala Ala Ile
Ser Ile Ser Thr Lys Val65 70 75
80 Leu Gly Leu Leu Gly Val Pro Phe Ala Ala Gln Ile Gly Gln Leu
Trp 85 90 95 Thr
Phe Ile Leu Asn Ala Leu Trp Pro Ser Asp Asn Thr Gln Trp Glu
100 105 110 Glu Phe Met Arg His
Val Glu Glu Leu Ile Asn Gln Arg Ile Ala Asp 115
120 125 Tyr Ala Arg Asn Lys Ala Leu Ala Glu
Leu Thr Gly Leu Gly Asn Asn 130 135
140 Leu Asp Leu Tyr Ile Glu Ala Leu Asp Asp Trp Lys Arg
Asn Pro Thr145 150 155
160 Ser Gln Glu Ala Lys Thr Arg Val Ile Asp Arg Phe Arg Ile Val Asp
165 170 175 Gly Leu Phe Glu
Ala Tyr Ile Pro Ser Phe Ala Val Ser Gly Tyr Gln 180
185 190 Val Gln Leu Leu Thr Val Tyr Ala Ala
Ala Ala Asn Leu His Leu Leu 195 200
205 Leu Leu Arg Asp Ser Thr Ile Tyr Gly Ile Asp Trp Gly Leu
Ser Gln 210 215 220
Thr Asn Val Asn Asp Asn Tyr Asn Arg Gln Ile Arg Leu Thr Ala Thr225
230 235 240 Tyr Ala Asn His Cys
Thr Thr Trp Tyr Gln Thr Gly Leu Glu Arg Leu 245
250 255 Arg Gly Ser Asn Ala Ser Ser Trp Val Thr
Tyr Asn Arg Phe Arg Arg 260 265
270 Glu Met Thr Leu Thr Val Leu Asp Ile Cys Ser Leu Phe Ser Asn
Tyr 275 280 285 Asp
Tyr Arg Ser Tyr Pro Ala Glu Val Arg Gly Glu Ile Thr Arg Glu 290
295 300 Ile Tyr Thr Asp Pro Val
Gly Val Gly Trp Val Asp Ser Ala Pro Ser305 310
315 320 Phe Gly Glu Ile Glu Asn Leu Ala Ile Arg Ala
Pro Arg Thr Val Thr 325 330
335 Trp Leu Asn Ser Thr Arg Ile Phe Thr Gly Arg Leu Gln Gly Trp Ser
340 345 350 Gly Thr Asn
Asn Tyr Trp Ala Ala His Met Gln Asn Phe Ser Glu Thr 355
360 365 Asn Ser Gly Asn Ile Gln Phe Glu
Gly Pro Leu Tyr Gly Ser Thr Val 370 375
380 Gly Thr Ile His Arg Thr Asp Asp Tyr Asp Met Gly Asn
Arg Asp Ile385 390 395
400 Tyr Thr Ile Thr Ser Gln Ala Val Leu Gly Leu Trp Ala Thr Gly Gln
405 410 415 Arg Val Leu Gly
Val Ala Ser Ala Arg Phe Thr Leu Arg Asn Leu Phe 420
425 430 Asn Asn Leu Thr Gln Val Leu Val Tyr
Glu Asn Pro Ile Ser Ser Thr 435 440
445 Phe Gly Ser Ser Thr Leu Thr His Glu Leu Ser Gly Glu Asn
Ser Asp 450 455 460
Arg Pro Thr Ser Ser Asp Tyr Ser His Arg Leu Thr Ser Ile Thr Gly465
470 475 480 Phe Arg Ala Gly Ala
Asn Gly Thr Val Pro Val Phe Gly Trp Thr Ser 485
490 495 Ala Thr Val Asp Arg Asn Asn Ile Ile Glu
Arg Asn Lys Ile Thr Gln 500 505
510 Phe Pro Gly Val Lys Ser His Thr Leu Asn Asn Cys Gln Val Val
Arg 515 520 525 Gly
Thr Gly Phe Thr Gly Gly Asp Trp Leu Arg Pro Asn Asn Asn Gly 530
535 540 Thr Phe Arg Leu Thr Ile
Thr Ser Phe Ser Ser Gln Ser Tyr Arg Ile545 550
555 560 Arg Leu Arg Tyr Ala Thr Ser Val Gly Asn Thr
Ser Leu Val Ile Ser 565 570
575 Ser Ser Asp Ala Gly Ile Ser Ser Thr Thr Ile Pro Leu Thr Ser Thr
580 585 590 Ile Thr Ser
Leu Pro Gln Thr Val Pro Tyr Gln Ala Phe Arg Val Val 595
600 605 Asp Leu Pro Ile Thr Phe Thr Thr
Pro Thr Thr Gln Arg Asn Tyr Thr 610 615
620 Phe Asp Phe Arg Leu Gln Asn Pro Ser Asn Ala Asn Val
Phe Ile Asp625 630 635
640 Arg Phe Glu Phe Val Pro Ile 645
481256PRTBrevibacillus laterosporus 48Met Asn Gln Asn Gln Asn Gln Asn Gln
Asn Lys Asn Glu Leu Gln Ile1 5 10
15 Ile Glu Pro Ser Ser Asp Ser Phe Leu Tyr Ser His Asn Asn
Tyr Pro 20 25 30
Tyr Ala Thr Asp Pro Asn Thr Val Leu Gln Gly Arg Asn Tyr Lys Glu 35
40 45 Trp Leu Asn Met Cys
Thr Gly Thr Asp Asp Ser Arg Ser Pro Glu Ala 50 55
60 Ala Ser Thr Ala Lys Ser Ala Ile Ser Val
Ala Ile Thr Ile Ser Thr65 70 75
80 Thr Ile Leu Gly Leu Leu Gly Val Pro Phe Ala Ser Gln Ile Gly
Ala 85 90 95 Phe
Tyr Asn Phe Val Leu Asn Thr Val Trp Pro Gln Gly Asn Asn Gln
100 105 110 Trp Glu Glu Phe Met
Arg His Val Glu Asp Leu Ile Asn Glu Arg Ile 115
120 125 Ala Asp Tyr Ala Arg Ser Lys Ala Leu
Ala Glu Leu Ala Gly Leu Gly 130 135
140 Asn Asn Leu Asp Leu Tyr Arg Glu Ala Phe Glu Asp Trp
Arg Arg Asn145 150 155
160 Pro Thr Ser Gln Gln Ala Lys Thr Arg Val Ile Glu Arg Phe Arg Ile
165 170 175 Leu Asp Gly Leu
Phe Glu Gln Tyr Met Pro Ser Phe Ala Val Gln Gly 180
185 190 Phe Gln Val Gln Leu Leu Thr Val Tyr
Ala Ser Ala Ala Asn Ile His 195 200
205 Leu Phe Leu Leu Arg Asp Ser Ser Ile Tyr Gly Leu Asp Trp
Gly Leu 210 215 220
Ser Gln Thr Asn Val Asn Glu Asn Tyr Asn Arg Gln Ile Arg His Ala225
230 235 240 Ala Thr Tyr Ala Asn
His Cys Thr Thr Trp Tyr Gln Thr Gly Leu Gln 245
250 255 Arg Leu Gln Gly Thr Asn Ala Thr Ser Trp
Val Ala Tyr Asn Arg Phe 260 265
270 Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Ser Ser Leu Phe
Ser 275 280 285 Asn
Tyr Asp Tyr Arg Ser Tyr Pro Thr Glu Val Arg Gly Glu Leu Thr 290
295 300 Arg Glu Ile Tyr Thr Asp
Pro Val Gly Arg Asn Trp Gln Asn Ser Ala305 310
315 320 Pro Ser Phe Ala Gln Ile Glu Asn Leu Ala Ile
Arg Ala Pro Arg Thr 325 330
335 Val Thr Trp Leu Asn Ser Thr Arg Ile Ser Thr Gly Thr Leu Gln Gly
340 345 350 Trp Ser Gly
Ser Asn Arg Tyr Trp Ala Ala His Met Gln Asn Phe Ser 355
360 365 Glu Thr Asn Ser Gly Asn Ile Arg
Phe Asp Gly Pro Leu Tyr Gly Ser 370 375
380 Thr Val Gly Thr Ile His Arg Thr Asp Asp Tyr Asp Met
Gly Asn Arg385 390 395
400 Asp Ile Tyr Thr Ile Thr Ser Glu Val Val Ala Ser Leu Trp Ala Thr
405 410 415 Gly Gln Thr Val
Leu Gly Val Ala Ser Ala Arg Phe Thr Leu Arg Asn 420
425 430 Leu Phe Asn Asn Leu Thr Gln Ala Leu
Val Tyr Glu Asn Pro Ile Ser 435 440
445 Ser Ser Phe Asn Arg Ser Thr Leu Thr His Glu Leu Pro Gly
Glu Asn 450 455 460
Ser Asp Arg Pro Thr Ser Ser Asp Tyr Ser His Arg Leu Ser Ser Ile465
470 475 480 Thr Gly Phe Arg Ala
Gly Ala Asn Gly Thr Val Pro Val Phe Gly Trp 485
490 495 Thr Ser Ala Thr Val Asp Arg Asn Asn Ile
Ile Glu Arg Asn Lys Ile 500 505
510 Thr Gln Phe Pro Gly Val Lys Ser His Thr Leu Asn Asn Cys Gln
Val 515 520 525 Val
Arg Gly Thr Gly Phe Thr Gly Gly Asp Trp Leu Arg Pro Asn Asn 530
535 540 Asn Gly Thr Phe Arg Leu
Thr Ile Thr Ser Phe Ser Ser Gln Ser Tyr545 550
555 560 Arg Ile Arg Leu Arg Tyr Ala Thr Ser Val Gly
Asn Thr Ser Leu Val 565 570
575 Ile Ser Ser Ser Asp Gly Gly Ile Ser Ser Thr Thr Ile Pro Leu Thr
580 585 590 Ser Thr Ile
Thr Ser Leu Pro Gln Thr Val Pro Tyr Gln Ala Phe Arg 595
600 605 Val Val Asp Leu Pro Ile Thr Phe
Thr Thr Pro Thr Thr Gln Arg Asn 610 615
620 Tyr Thr Phe Asp Phe Arg Leu Gln Asn Pro Ser Asn Ala
Asn Val Phe625 630 635
640 Ile Asp Arg Ile Glu Phe Val Pro Ile Gly Gly Ser Leu Ser Glu Tyr
645 650 655 Glu Thr Lys His
Gln Leu Glu Lys Ala Arg Lys Ala Val Asn Asp Leu 660
665 670 Phe Thr Asn Glu Ser Lys Asn Val Leu
Lys Lys Asp Thr Thr Asp Tyr 675 680
685 Asp Ile Asp Gln Ala Ala Asn Leu Val Glu Cys Val Ser Asp
Glu Cys 690 695 700
Ala Asn Ala Lys Met Ile Leu Leu Asp Glu Val Lys Tyr Ala Lys Gln705
710 715 720 Leu Ser Glu Ala Arg
Asn Leu Leu Leu Asn Gly Asn Phe Asp Asn Ile 725
730 735 Asp Arg Asp Gly Glu Asn Pro Trp Lys Thr
Ser Pro Asn Val Thr Ile 740 745
750 Gln Glu Asn Asn Pro Ile Phe Lys Gly Arg Tyr Leu Ser Met Ser
Gly 755 760 765 Ala
Asn Asn Ile Glu Ala Thr Asn Glu Ile Phe Pro Thr Tyr Ala Tyr 770
775 780 Gln Lys Ile Asp Glu Ala
Lys Leu Lys Pro Tyr Thr Arg Tyr Lys Val785 790
795 800 Arg Gly Phe Val Gly Asn Ser Lys Asp Leu Glu
Leu Leu Val Thr Arg 805 810
815 Tyr Asp Glu Glu Val Asp Ala Ile Leu Asn Val Pro Asn Asp Ile Pro
820 825 830 His Ala Pro
Pro Pro Phe Cys Gly Glu Phe Asp Arg Cys Asn Pro His 835
840 845 Ser Tyr Pro Pro Met Asn Pro Glu
Cys His His Asp Val Ile Asn Asn 850 855
860 Ile Glu Ile Ser Ser Pro Cys Gln His Asn Lys Met Val
Asp Asn Ala865 870 875
880 Asp Ile Ser Tyr Arg His Ser His Lys Lys His Gly Ile Cys His Glu
885 890 895 Ser His His Phe
Glu Phe His Ile Asp Thr Gly Lys Ile Asp Leu Val 900
905 910 Glu Asn Leu Gly Ile Trp Val Ile Phe
Lys Ile Cys Ser Thr Asp Gly 915 920
925 Tyr Ala Thr Leu Asp Asn Leu Glu Val Ile Glu Glu Arg Pro
Leu Gly 930 935 940
Ala Glu Ser Leu Glu Arg Val Lys Arg Arg Glu Lys Lys Trp Lys His945
950 955 960 His Met Glu His Lys
Cys Ser Glu Thr Lys Leu Ala Tyr His Ala Ala 965
970 975 Lys Gln Ala Leu Val Gly Leu Phe Thr Asn
Thr Glu Tyr Asp Arg Leu 980 985
990 Lys Phe Glu Thr Thr Ile Ser Asn Ile Leu Phe Ala Asp Tyr Leu
Val 995 1000 1005 Gln
Ser Ile Pro Tyr Val Tyr Asn Lys Trp Leu Pro Asp Val Pro Gly 1010
1015 1020 Met Asn Phe Glu Ile Tyr
Thr Glu Leu Lys Asn Leu Tyr Thr Gly Ala1025 1030
1035 1040 Phe Asn Leu Tyr Asp Gln Arg Asn Ile Ile Lys
Asn Gly Asp Phe Asn 1045 1050
1055 Arg Gly Leu Met His Trp His Ala Thr Pro His Ala Arg Val Glu Gln
1060 1065 1070 Ile Asp Asn
Arg Ser Val Leu Val Leu Pro Asn Tyr Ala Ala Asn Val 1075
1080 1085 Ser Gln Glu Val Cys Leu Glu His
Asn Arg Gly Tyr Val Leu Arg Val 1090 1095
1100 Thr Ala Lys Lys Glu Gly Pro Gly Ile Gly Tyr Ile Thr
Phe Ser Asp1105 1110 1115
1120 Cys Ala Asn Asn Ile Glu Lys Leu Thr Phe Thr Ser Cys Asp Tyr Gly
1125 1130 1135 Thr Asn Glu Val
Pro Tyr Glu Gln Ser Asn Tyr Pro Thr Asp Gly Val 1140
1145 1150 Ser Tyr Gly His His Gly Cys Asn Ile
Asp Arg Val Arg Tyr Glu Glu 1155 1160
1165 Ser Gly Tyr Arg Thr Asp Gly Val Pro Tyr Glu Gln Ser Gly
Tyr Arg 1170 1175 1180
Ala Asp Gly Val Ser Tyr Glu Gln His Gly Cys His Thr Asp Gly Val1185
1190 1195 1200 Pro Tyr Lys Gln His
Gly Cys Arg Thr Asp Arg Ser Arg Asp Glu Gln 1205
1210 1215 Leu Gly Tyr Val Thr Lys Thr Ile Asp Val
Phe Pro Asp Thr Asp Lys 1220 1225
1230 Val Arg Ile Asp Ile Gly Glu Thr Glu Gly Thr Phe Lys Val Glu
Ser 1235 1240 1245 Val
Glu Leu Ile Cys Met Glu Glu 1250 1255
49649PRTBrevibacillus laterosporus 49Met Asn Gln Asn Gln Asn Gln Asn Gln
Asn Lys Asn Glu Leu Gln Ile1 5 10
15 Ile Glu Pro Ser Ser Asp Ser Phe Leu Tyr Ser His Asn Asn
Tyr Pro 20 25 30
Tyr Ala Thr Asp Pro Asn Thr Val Leu Gln Gly Arg Asn Tyr Lys Glu 35
40 45 Trp Leu Asn Met Cys
Thr Gly Thr Asp Asp Ser Arg Ser Pro Glu Ala 50 55
60 Ala Ser Thr Ala Lys Ser Ala Ile Ser Val
Ala Ile Thr Ile Ser Thr65 70 75
80 Thr Ile Leu Gly Leu Leu Gly Val Pro Phe Ala Ser Gln Ile Gly
Ala 85 90 95 Phe
Tyr Asn Phe Val Leu Asn Thr Val Trp Pro Gln Gly Asn Asn Gln
100 105 110 Trp Glu Glu Phe Met
Arg His Val Glu Asp Leu Ile Asn Glu Arg Ile 115
120 125 Ala Asp Tyr Ala Arg Ser Lys Ala Leu
Ala Glu Leu Ala Gly Leu Gly 130 135
140 Asn Asn Leu Asp Leu Tyr Arg Glu Ala Phe Glu Asp Trp
Arg Arg Asn145 150 155
160 Pro Thr Ser Gln Gln Ala Lys Thr Arg Val Ile Glu Arg Phe Arg Ile
165 170 175 Leu Asp Gly Leu
Phe Glu Gln Tyr Met Pro Ser Phe Ala Val Gln Gly 180
185 190 Phe Gln Val Gln Leu Leu Thr Val Tyr
Ala Ser Ala Ala Asn Ile His 195 200
205 Leu Phe Leu Leu Arg Asp Ser Ser Ile Tyr Gly Leu Asp Trp
Gly Leu 210 215 220
Ser Gln Thr Asn Val Asn Glu Asn Tyr Asn Arg Gln Ile Arg His Ala225
230 235 240 Ala Thr Tyr Ala Asn
His Cys Thr Thr Trp Tyr Gln Thr Gly Leu Gln 245
250 255 Arg Leu Gln Gly Thr Asn Ala Thr Ser Trp
Val Ala Tyr Asn Arg Phe 260 265
270 Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Ser Ser Leu Phe
Ser 275 280 285 Asn
Tyr Asp Tyr Arg Ser Tyr Pro Thr Glu Val Arg Gly Glu Leu Thr 290
295 300 Arg Glu Ile Tyr Thr Asp
Pro Val Gly Arg Asn Trp Gln Asn Ser Ala305 310
315 320 Pro Ser Phe Ala Gln Ile Glu Asn Leu Ala Ile
Arg Ala Pro Arg Thr 325 330
335 Val Thr Trp Leu Asn Ser Thr Arg Ile Ser Thr Gly Thr Leu Gln Gly
340 345 350 Trp Ser Gly
Ser Asn Arg Tyr Trp Ala Ala His Met Gln Asn Phe Ser 355
360 365 Glu Thr Asn Ser Gly Asn Ile Arg
Phe Asp Gly Pro Leu Tyr Gly Ser 370 375
380 Thr Val Gly Thr Ile His Arg Thr Asp Asp Tyr Asp Met
Gly Asn Arg385 390 395
400 Asp Ile Tyr Thr Ile Thr Ser Glu Val Val Ala Ser Leu Trp Ala Thr
405 410 415 Gly Gln Thr Val
Leu Gly Val Ala Ser Ala Arg Phe Thr Leu Arg Asn 420
425 430 Leu Phe Asn Asn Leu Thr Gln Ala Leu
Val Tyr Glu Asn Pro Ile Ser 435 440
445 Ser Ser Phe Asn Arg Ser Thr Leu Thr His Glu Leu Pro Gly
Glu Asn 450 455 460
Ser Asp Arg Pro Thr Ser Ser Asp Tyr Ser His Arg Leu Ser Ser Ile465
470 475 480 Thr Gly Phe Arg Ala
Gly Ala Asn Gly Thr Val Pro Val Phe Gly Trp 485
490 495 Thr Ser Ala Thr Val Asp Arg Asn Asn Ile
Ile Glu Arg Asn Lys Ile 500 505
510 Thr Gln Phe Pro Gly Val Lys Ser His Thr Leu Asn Asn Cys Gln
Val 515 520 525 Val
Arg Gly Thr Gly Phe Thr Gly Gly Asp Trp Leu Arg Pro Asn Asn 530
535 540 Asn Gly Thr Phe Arg Leu
Thr Ile Thr Ser Phe Ser Ser Gln Ser Tyr545 550
555 560 Arg Ile Arg Leu Arg Tyr Ala Thr Ser Val Gly
Asn Thr Ser Leu Val 565 570
575 Ile Ser Ser Ser Asp Gly Gly Ile Ser Ser Thr Thr Ile Pro Leu Thr
580 585 590 Ser Thr Ile
Thr Ser Leu Pro Gln Thr Val Pro Tyr Gln Ala Phe Arg 595
600 605 Val Val Asp Leu Pro Ile Thr Phe
Thr Thr Pro Thr Thr Gln Arg Asn 610 615
620 Tyr Thr Phe Asp Phe Arg Leu Gln Asn Pro Ser Asn Ala
Asn Val Phe625 630 635
640 Ile Asp Arg Ile Glu Phe Val Pro Ile 645
50317PRTBrevibacillus laterosporus 50Met Lys Lys Phe Ala Ser Leu Ile Leu
Ile Ser Val Phe Leu Phe Ser1 5 10
15 Ser Thr Gln Phe Val His Ala Ser Ser Thr Asp Val Gln Glu
Arg Leu 20 25 30
Arg Asp Leu Ala Arg Glu Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp 35
40 45 Asn Thr Asn Phe Lys
Pro Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro 50 55
60 Thr Glu Gly Ile Val Phe Leu Thr Pro Pro
Lys Asn Val Ile Gly Glu65 70 75
80 Arg Arg Ile Ser Gln Tyr Lys Val Asn Asn Ala Trp Ala Thr Leu
Glu 85 90 95 Gly
Ser Pro Thr Glu Val Ser Gly Thr Pro Leu Tyr Val Gly Lys Asn
100 105 110 Val Leu Asp Asn Ser
Lys Gly Thr Ser Asp Gln Glu Leu Leu Thr Pro 115
120 125 Glu Phe Asn Tyr Thr Tyr Thr Glu Ser
Thr Ser Asn Thr Thr Thr His 130 135
140 Gly Leu Lys Leu Gly Val Lys Thr Thr Ala Thr Met Lys
Phe Pro Ile145 150 155
160 Ala Gln Gly Ser Met Glu Ala Ser Thr Glu Tyr Asn Phe Gln Asp Ser
165 170 175 Ser Thr Asp Thr
Thr Thr Lys Thr Val Ser Tyr Lys Ser Pro Ser Gln 180
185 190 Lys Ile Lys Val Pro Ala Gly Lys Thr
Phe Arg Val Leu Ala Tyr Leu 195 200
205 Asn Thr Gly Ser Ile Ser Gly Glu Ala Asn Leu Tyr Ala Asn
Val Gly 210 215 220
Gly Ile Ala Trp Gly Val Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val225
230 235 240 Asn Ile Gly Ala Val
Leu Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp 245
250 255 Phe Arg Asn Phe Gln Pro Ser Gly Arg Asp
Val Ile Val Lys Gly Gln 260 265
270 Gly Thr Phe Lys Ser Asn Tyr Gly Thr Asp Phe Ile Leu Lys Ile
Glu 275 280 285 Asp
Ile Thr Asp Ser Lys Leu Arg Asn Asn Asn Gly Ser Gly Thr Val 290
295 300 Val Gln Glu Ile Lys Val
Pro Leu Ile Arg Thr Glu Ile305 310 315
511154PRTBacillus thuringiensis 51Met Asn Phe Leu Phe Leu Val Asn
Tyr Glu Lys Asn Lys Phe Lys Tyr1 5 10
15 Asn Ile Gln Gly Asp Leu Asn Met Asn Gln Lys Asn Tyr
Asp Ile Ile 20 25 30
Gly Ser Ser Thr Asn Gly Thr Thr Lys Leu Pro Glu Asp Tyr Asn Ile
35 40 45 Ile Ile Ser Pro
Asp Ala Ala Pro Glu Ala Val Thr Ile Ala Ile Ser 50 55
60 Ile Thr Gly Glu Val Leu Ser Leu Phe
Gly Val Pro Gly Ala Thr Leu65 70 75
80 Gly Ser Thr Leu Leu Asn Thr Leu Val Asp Lys Leu Trp Pro
Thr Asn 85 90 95
Thr Asn Thr Val Trp Gly Thr Phe Thr Glu Glu Thr Ala Lys Leu Ile
100 105 110 Asn Glu Val Tyr Asn
Pro Ser Asp Pro Val Val Lys Asp Ala Asp Ala 115
120 125 Arg Leu Thr Ser Leu His Glu Ser Leu
Lys Leu Tyr Gln Leu Ala Phe 130 135
140 Gly Asn Trp Phe Lys Ser Gln Asp Asn Ser Lys Leu Lys
Glu Glu Val145 150 155
160 Arg Arg Gln Phe Asp Ile Thr His Asn Arg Phe Val Thr Ser Met Pro
165 170 175 Phe Phe Lys Val
Ser Asp Tyr Glu Ile Arg Leu Leu Thr Asn Tyr Ala 180
185 190 Gln Ala Ala Asn Leu His Leu Thr Phe
Leu Arg Asp Ala Ser Ile Tyr 195 200
205 Gly Leu Asp Trp Gly Phe Ser Asp Glu His Ser Asn Asp Leu
Tyr Glu 210 215 220
Gln Gln Lys Asn Arg Thr Gly Glu Tyr Thr Asp His Cys Val Lys Trp225
230 235 240 Tyr Asn Ala Gly Leu
Glu Lys Leu Lys Gly Asn Leu Thr Gly Glu Asn 245
250 255 Trp Tyr Thr Tyr Asn Arg Phe Arg Arg Glu
Met Thr Leu Met Val Leu 260 265
270 Asp Val Val Ala Leu Phe Pro Asn Tyr Asp Thr Arg Met Tyr Pro
Ile 275 280 285 Ala
Thr Ser Ser Glu Leu Thr Arg Met Ile Tyr Thr Asp Pro Ile Ala 290
295 300 Tyr Thr Gln Ser Asp Pro
Trp Tyr Lys Ile Thr Ser Leu Ser Phe Ser305 310
315 320 Asn Ile Glu Asn Ser Ala Ile Pro Ser Pro Ser
Phe Phe Arg Trp Leu 325 330
335 Lys Ser Val Ser Ile Asn Ser Gln Trp Trp Gly Ser Gly Pro Asn Gln
340 345 350 Thr Tyr Tyr
Trp Val Gly His Glu Leu Val Tyr Ser Asn Ser Asn Tyr 355
360 365 Asn Gln Ser Leu Lys Val Lys Tyr
Gly Asp Pro Asn Ser Tyr Ile Glu 370 375
380 Pro Pro Asp Ser Phe Ser Phe Ser Ser Thr Asp Val Tyr
Arg Thr Ile385 390 395
400 Ser Val Val Arg Asn Ser Ile Ser Asn Tyr Ile Val Ser Glu Val Gln
405 410 415 Phe Asn Ser Ile
Ser Asn Thr Asn Gln Ile Ser Glu Glu Ile Tyr Lys 420
425 430 His Gln Ser Asn Trp Asn Arg Arg Glu
Thr Lys Asp Ser Ile Thr Glu 435 440
445 Leu Ser Leu Ala Ala Asn Pro Pro Thr Thr Phe Gly Asn Val
Ala Glu 450 455 460
Tyr Ser His Arg Leu Ala Tyr Ile Ser Glu Ala Tyr Gln Ser Asn Asn465
470 475 480 Pro Ser Lys Tyr Pro
Ala Tyr Ile Pro Val Phe Gly Trp Thr His Thr 485
490 495 Ser Val Arg Tyr Asp Asn Lys Ile Phe Pro
Asp Lys Ile Thr Gln Ile 500 505
510 Pro Ala Val Lys Ser Ser Ser Ala Glu Gly Gly Thr Trp Lys Asn
Ile 515 520 525 Ala
Lys Gly Pro Gly Phe Thr Gly Gly Asp Val Thr Thr Ala Val Ser 530
535 540 Pro Ala Phe Ile Thr Asp
Met Ile Lys Ile His Val Thr Leu Asp Pro545 550
555 560 Asn Ser Leu Ser Gln Lys Tyr Arg Ala Arg Leu
Arg Tyr Ala Ser Asn 565 570
575 Ala Tyr Val Ala Ala Thr Leu Tyr Thr Asn Ser Ser Ser Asn Tyr Asn
580 585 590 Phe Glu Leu
Thr Lys Gly Thr Thr Glu Gln Phe Thr Thr Tyr Asn Ser 595
600 605 Tyr Gln Tyr Val Asp Ile Pro Gly
Ser Ile Gln Phe Asn Thr Thr Ser 610 615
620 Asp Thr Val Ser Val Tyr Leu His Met Asp Ser Thr Thr
Asn Ala Asn625 630 635
640 Val His Val Asp Arg Ile Glu Phe Ile Pro Val Asp Glu Asn Tyr Asp
645 650 655 Asn Arg Val Thr
Leu Glu Lys Ala Gln Lys Ala Val Asn Ala Leu Phe 660
665 670 Thr Ala Gly Arg His Ala Leu Gln Thr
Asp Val Thr Asp Phe Lys Val 675 680
685 Asp Gln Val Ser Ile Leu Val Asp Cys Val Ser Gly Glu Leu
Tyr Pro 690 695 700
Asn Glu Lys Arg Glu Leu Leu Ser Leu Val Lys Tyr Ala Lys Arg Leu705
710 715 720 Ser Tyr Ser Arg Asn
Leu Leu Leu Asp Pro Thr Phe Asp Ser Ile Asn 725
730 735 Ser Ser Glu Glu Asn Gly Trp His Gly Ser
Asn Gly Ile Ala Ile Gly 740 745
750 Asn Gly Asn Phe Val Phe Lys Gly Asn Tyr Leu Ile Phe Ser Gly
Thr 755 760 765 Asn
Asp Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu Ser 770
775 780 Lys Leu Lys Glu Tyr Thr
Arg Tyr Lys Leu Arg Gly Phe Ile Glu Ser785 790
795 800 Ser Gln Asp Leu Glu Ala Tyr Val Ile Arg Tyr
Asp Ala Lys Tyr Glu 805 810
815 Thr Leu Asp Val Ser Asn Asn Leu Tyr Pro Asp Ile Ser Pro Val Asn
820 825 830 Ala Cys Gly
Glu Pro Asn Arg Cys Ala Ala Leu Pro Tyr Leu Asp Glu 835
840 845 Asn Pro Arg Leu Glu Cys Ser Ser
Ile Gln Asp Gly Ile Leu Ser Asp 850 855
860 Ser His Ser Phe Ser Leu Asn Ile Asp Thr Gly Ser Ile
Asp Ser Asn865 870 875
880 Glu Asn Val Gly Ile Trp Val Leu Phe Lys Ile Ser Thr Pro Glu Gly
885 890 895 Tyr Ala Lys Phe
Gly Asn Leu Glu Val Ile Glu Asp Gly Pro Val Ile 900
905 910 Gly Glu Ala Leu Ala Arg Val Lys Arg
Gln Glu Thr Lys Trp Arg Asn 915 920
925 Lys Leu Thr Gln Leu Arg Thr Glu Thr Gln Ala Ile Tyr Thr
Arg Ala 930 935 940
Lys Gln Ala Ile Asp Asn Leu Phe Thr Asn Ala Gln Asp Ser His Leu945
950 955 960 Lys Ile Gly Ala Thr
Phe Ala Ser Ile Val Ala Ala Arg Lys Ile Val 965
970 975 Gln Ser Ile Arg Glu Ala Tyr Met Pro Trp
Leu Ser Ile Val Pro Gly 980 985
990 Val Asn Tyr Pro Ile Phe Thr Glu Leu Asn Glu Arg Val Gln Gln
Ala 995 1000 1005 Phe
Gln Leu Tyr Asp Val Arg Asn Val Val Arg Asn Gly Arg Phe Leu 1010
1015 1020 Asn Gly Val Ser Asp Trp
Ile Val Thr Ser Asp Val Thr Val Gln Glu1025 1030
1035 1040 Glu Asn Gly Asn Asn Val Leu Val Leu Ser Asn
Trp Asp Ala Gln Val 1045 1050
1055 Leu Gln Cys Leu Lys Leu Tyr Gln Asp Arg Gly Tyr Ile Leu Arg Val
1060 1065 1070 Thr Ala Arg
Lys Glu Gly Leu Gly Glu Gly Tyr Ile Thr Ile Thr Asp 1075
1080 1085 Glu Glu Gly Tyr Thr Asp Gln Leu
Thr Phe Gly Thr Cys Glu Glu Ile 1090 1095
1100 Asp Ala Ser Asn Thr Phe Val Ser Thr Gly Tyr Ile Thr
Lys Glu Leu1105 1110 1115
1120 Glu Phe Phe Pro Asp Thr Glu Lys Val Arg Ile Glu Val Gly Glu Thr
1125 1130 1135 Glu Gly Thr Phe
Arg Val Glu Ser Val Glu Leu Phe Leu Met Glu Glu 1140
1145 1150 His Cys52651PRTBacillus
thuringiensis 52Met Asn Phe Leu Phe Leu Val Asn Tyr Glu Lys Asn Lys Phe
Lys Tyr1 5 10 15
Asn Ile Gln Gly Asp Leu Asn Met Asn Gln Lys Asn Tyr Asp Ile Ile
20 25 30 Gly Ser Ser Thr Asn
Gly Thr Thr Lys Leu Pro Glu Asp Tyr Asn Ile 35 40
45 Ile Ile Ser Pro Asp Ala Ala Pro Glu Ala
Val Thr Ile Ala Ile Ser 50 55 60
Ile Thr Gly Glu Val Leu Ser Leu Phe Gly Val Pro Gly Ala Thr
Leu65 70 75 80 Gly
Ser Thr Leu Leu Asn Thr Leu Val Asp Lys Leu Trp Pro Thr Asn
85 90 95 Thr Asn Thr Val Trp Gly
Thr Phe Thr Glu Glu Thr Ala Lys Leu Ile 100
105 110 Asn Glu Val Tyr Asn Pro Ser Asp Pro Val
Val Lys Asp Ala Asp Ala 115 120
125 Arg Leu Thr Ser Leu His Glu Ser Leu Lys Leu Tyr Gln Leu
Ala Phe 130 135 140
Gly Asn Trp Phe Lys Ser Gln Asp Asn Ser Lys Leu Lys Glu Glu Val145
150 155 160 Arg Arg Gln Phe Asp
Ile Thr His Asn Arg Phe Val Thr Ser Met Pro 165
170 175 Phe Phe Lys Val Ser Asp Tyr Glu Ile Arg
Leu Leu Thr Asn Tyr Ala 180 185
190 Gln Ala Ala Asn Leu His Leu Thr Phe Leu Arg Asp Ala Ser Ile
Tyr 195 200 205 Gly
Leu Asp Trp Gly Phe Ser Asp Glu His Ser Asn Asp Leu Tyr Glu 210
215 220 Gln Gln Lys Asn Arg Thr
Gly Glu Tyr Thr Asp His Cys Val Lys Trp225 230
235 240 Tyr Asn Ala Gly Leu Glu Lys Leu Lys Gly Asn
Leu Thr Gly Glu Asn 245 250
255 Trp Tyr Thr Tyr Asn Arg Phe Arg Arg Glu Met Thr Leu Met Val Leu
260 265 270 Asp Val Val
Ala Leu Phe Pro Asn Tyr Asp Thr Arg Met Tyr Pro Ile 275
280 285 Ala Thr Ser Ser Glu Leu Thr Arg
Met Ile Tyr Thr Asp Pro Ile Ala 290 295
300 Tyr Thr Gln Ser Asp Pro Trp Tyr Lys Ile Thr Ser Leu
Ser Phe Ser305 310 315
320 Asn Ile Glu Asn Ser Ala Ile Pro Ser Pro Ser Phe Phe Arg Trp Leu
325 330 335 Lys Ser Val Ser
Ile Asn Ser Gln Trp Trp Gly Ser Gly Pro Asn Gln 340
345 350 Thr Tyr Tyr Trp Val Gly His Glu Leu
Val Tyr Ser Asn Ser Asn Tyr 355 360
365 Asn Gln Ser Leu Lys Val Lys Tyr Gly Asp Pro Asn Ser Tyr
Ile Glu 370 375 380
Pro Pro Asp Ser Phe Ser Phe Ser Ser Thr Asp Val Tyr Arg Thr Ile385
390 395 400 Ser Val Val Arg Asn
Ser Ile Ser Asn Tyr Ile Val Ser Glu Val Gln 405
410 415 Phe Asn Ser Ile Ser Asn Thr Asn Gln Ile
Ser Glu Glu Ile Tyr Lys 420 425
430 His Gln Ser Asn Trp Asn Arg Arg Glu Thr Lys Asp Ser Ile Thr
Glu 435 440 445 Leu
Ser Leu Ala Ala Asn Pro Pro Thr Thr Phe Gly Asn Val Ala Glu 450
455 460 Tyr Ser His Arg Leu Ala
Tyr Ile Ser Glu Ala Tyr Gln Ser Asn Asn465 470
475 480 Pro Ser Lys Tyr Pro Ala Tyr Ile Pro Val Phe
Gly Trp Thr His Thr 485 490
495 Ser Val Arg Tyr Asp Asn Lys Ile Phe Pro Asp Lys Ile Thr Gln Ile
500 505 510 Pro Ala Val
Lys Ser Ser Ser Ala Glu Gly Gly Thr Trp Lys Asn Ile 515
520 525 Ala Lys Gly Pro Gly Phe Thr Gly
Gly Asp Val Thr Thr Ala Val Ser 530 535
540 Pro Ala Phe Ile Thr Asp Met Ile Lys Ile His Val Thr
Leu Asp Pro545 550 555
560 Asn Ser Leu Ser Gln Lys Tyr Arg Ala Arg Leu Arg Tyr Ala Ser Asn
565 570 575 Ala Tyr Val Ala
Ala Thr Leu Tyr Thr Asn Ser Ser Ser Asn Tyr Asn 580
585 590 Phe Glu Leu Thr Lys Gly Thr Thr Glu
Gln Phe Thr Thr Tyr Asn Ser 595 600
605 Tyr Gln Tyr Val Asp Ile Pro Gly Ser Ile Gln Phe Asn Thr
Thr Ser 610 615 620
Asp Thr Val Ser Val Tyr Leu His Met Asp Ser Thr Thr Asn Ala Asn625
630 635 640 Val His Val Asp Arg
Ile Glu Phe Ile Pro Val 645 650
531206PRTBacillus thuringiensis 53Met Arg Leu Lys Lys Leu Leu Val Cys Asn
Ile Arg Ile Gly Gly Thr1 5 10
15 Asn Met Asn Leu Gly Asn Tyr Asn Glu Phe Asp Ile Ile Asp Ile
Thr 20 25 30 Glu
Asn Asn Gln Thr Lys Thr Ser Arg Tyr Asn Asn Val Asn Arg Gln 35
40 45 Glu Asn Pro Ser Asn Met
Ile Ile Ser Asn Pro Ser Ser Asn Tyr Pro 50 55
60 Leu Ala Asn Asn Pro Asn Thr Pro Phe Gln Asn
Ile Asn Tyr Lys Asp65 70 75
80 Phe Leu Asn Met Asn Glu Glu Ile Ala Pro Tyr Ala Ser Ser Lys Asp
85 90 95 Val Ile Phe
Ser Ser Met Asn Ile Ile Arg Thr Phe Met Gly Phe Ala 100
105 110 Gly His Gly Thr Ala Gly Gly Ile
Val Ala Leu Phe Thr Glu Val Leu 115 120
125 Arg Leu Leu Trp Pro Asn Lys Gln Asp Glu Leu Trp Glu
Ser Phe Met 130 135 140
Lys Glu Val Glu Lys Leu Ile Glu Gln Glu Ile Thr Asp Ala Val Val145
150 155 160 Ser Lys Ala Leu Ala
Glu Leu Glu Gly Leu Arg Asn Ala Leu Gln Gly 165
170 175 Tyr Thr Asp Ala Leu Glu Ala Trp Gln Asn
Asn Arg Ser Asp Lys Leu 180 185
190 Lys Gln Leu Leu Val Tyr Asp Arg Phe Val Ser Thr Glu Asn Leu
Phe 195 200 205 Lys
Phe Ala Met Pro Ser Phe Arg Val Gly Gly Phe Glu Val Pro Leu 210
215 220 Leu Thr Val Tyr Ala Gln
Ala Ala Asn Leu His Leu Leu Leu Leu Lys225 230
235 240 Asn Ser Glu Leu Phe Gly Ala Glu Trp Gly Met
Gln Gln Tyr Glu Ile 245 250
255 Asp Leu Phe Tyr Asn Glu Gln Lys Asp Tyr Val Val Glu Tyr Thr Asp
260 265 270 His Cys Val
Lys Trp Tyr Thr Glu Gly Leu Asn Arg Leu Lys Asn Ala 275
280 285 Ser Gly Val Lys Gly Lys Val Trp
Glu Glu Tyr Asn Arg Phe Arg Arg 290 295
300 Glu Met Thr Ile Met Val Leu Asp Leu Leu Pro Leu Phe
Pro Ile Tyr305 310 315
320 Asp Val Arg Thr Tyr Pro Thr Glu Thr Val Thr Glu Leu Thr Arg Gln
325 330 335 Ile Phe Thr Asp
Pro Ile Gly Leu Arg Gly Ile Asn Glu Ser Lys Tyr 340
345 350 Pro Asp Trp Tyr Gly Ala Ala Ser Asp
Ser Phe Ser Leu Ile Glu Asn 355 360
365 Arg Ala Val Pro Gln Pro Ser Leu Phe Gln Trp Leu Thr Glu
Phe Lys 370 375 380
Val Tyr Thr Lys Tyr Val Glu Pro Asn Asp Lys Leu Thr Ile Leu Ala385
390 395 400 Gly His Ser Val Thr
Thr Gln Tyr Thr Ser Tyr Tyr Lys Lys Ser Thr 405
410 415 Phe Thr Tyr Gly Asp Thr Ser Ser Ala Asn
Ser Ser Arg Thr Phe Asp 420 425
430 Leu Leu Ala Lys Asp Val Tyr Gln Val Asp Ser Val Ala Ala Ala
Ser 435 440 445 Lys
Ser Ala Thr Trp Tyr Leu Ala Val Pro Glu Met Arg Leu Tyr Ser 450
455 460 Ile Asn Thr Asn Asn Ile
Leu Ser Glu Asp Tyr Phe Ser Leu Ser Thr465 470
475 480 Asn Ile Pro Ser Ser Lys Met Arg Arg Met Tyr
Ser Ser Glu Glu Leu 485 490
495 Pro Ile Gly Ile Ser Asp Thr Pro Ile Tyr Gly Asp Leu Glu Glu Tyr
500 505 510 Ser His Arg
Leu Ser Phe Ile Ser Glu Ile Met His Asn Ser Gly Ser 515
520 525 Val Thr Gly Ser Asn Asn Ile Lys
Gly Ile Ile Pro Val Leu Gly Trp 530 535
540 Thr His Thr Ser Val Ser Pro Glu Asn Tyr Ile His Arg
Asp Lys Ile545 550 555
560 Ser Gln Leu Tyr Ala Val Lys Ala Tyr Thr Thr Ser Asn Val Ser Val
565 570 575 Val Gly Gly Pro
Gly Phe Leu Gly Gly Asn Ile Ile Lys Gly Asn Asn 580
585 590 Asp Pro Ala Ser Tyr Thr Gly Ser Val
Ser Trp Ala Ile Arg Leu Asp 595 600
605 Gly Ser Thr Val Ser Arg Phe Arg Leu Arg Ile Pro Tyr Ala
Ala Glu 610 615 620
Thr Asp Gly Thr Phe Ser Ile Thr Val Arg Asp Asp Leu Gly Pro Phe625
630 635 640 Thr Ile Lys Lys Asp
Phe Ile Ala Thr Met Lys Pro Gly Asp Pro Leu 645
650 655 Ser Tyr Gly Lys Phe Glu Tyr Leu Glu Phe
Glu Gln Thr Met Ser Leu 660 665
670 Asn Asn Lys His Gly Gln Phe Phe Val His Thr Glu Asn Leu Lys
Asp 675 680 685 Arg
Asn Ser Ser Val Tyr Trp Asn Arg Val Glu Ile Ile Pro Val Asp 690
695 700 Glu Asn Tyr Asp Asn Arg
Val Arg Leu Glu Lys Ala Gln Lys Ala Val705 710
715 720 Asn Ala Leu Phe Thr Ala Gly Arg His Ala Leu
Gln Thr Asn Val Thr 725 730
735 Asp Tyr Lys Val Asp Gln Val Ser Ile Leu Val Asp Ser Val Ser Gly
740 745 750 Glu Leu Tyr
Pro Asn Glu Lys Arg Glu Leu Gln Ser Leu Val Lys Tyr 755
760 765 Ala Lys Arg Leu Ser Tyr Ser Arg
Asn Leu Leu Leu Asp Pro Thr Phe 770 775
780 Asp Ser Ile Asn Ser Ser Glu Glu Asn Gly Trp Tyr Gly
Ser Asn Gly785 790 795
800 Ile Ala Ile Gly Asn Gly Asn Phe Val Phe Lys Gly Asn Tyr Leu Asn
805 810 815 Phe Ser Gly Thr
Asn Asp Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln Lys 820
825 830 Ile Asp Glu Ser Lys Leu Lys Glu Tyr
Thr Arg Tyr Lys Leu Arg Gly 835 840
845 Phe Ile Glu Ser Ser Gln Asp Leu Glu Ala Tyr Val Val Arg
Tyr Asp 850 855 860
Ala Lys His Glu Thr Leu Asp Val Ser Asn Asn Leu Phe Pro Asp Ile865
870 875 880 Ser Pro Val Asn Ala
Cys Gly Glu Pro Asn Arg Cys Ala Ala Leu Pro 885
890 895 Tyr Leu Asp Lys Asn Pro Arg Leu Glu Cys
Ser Leu Ile Gln Asp Gly 900 905
910 Ile Leu Ser Asp Ser His Ser Phe Ser Leu Asn Ile Asp Thr Gly
Ser 915 920 925 Ile
Asp Ser Thr Glu Asn Val Gly Ile Trp Val Leu Phe Lys Ile Ser 930
935 940 Thr Pro Glu Gly Tyr Ala
Lys Phe Gly Asn Leu Glu Val Ile Glu Tyr945 950
955 960 Gly Pro Val Ile Gly Glu Ala Leu Ala Arg Val
Lys Arg Gln Glu Thr 965 970
975 Lys Trp Arg Asn Lys Leu Thr Gln Leu Arg Thr Glu Thr Gln Ala Ile
980 985 990 Tyr Thr Arg
Ala Lys Gln Ala Ile Asp Asn Leu Phe Thr Asn Thr Gln 995
1000 1005 Asp Ser Tyr Leu Lys Ile Gly Ala
Thr Phe Ala Ser Ile Val Ala Ala 1010 1015
1020 Arg Lys Ile Val Gln Ser Ile Arg Glu Ala Tyr Met Ser
Trp Leu Ser1025 1030 1035
1040 Ile Val Pro Gly Val Asn Tyr Pro Ile Phe Thr Glu Leu Asn Glu Arg
1045 1050 1055 Val Gln Arg Ala
Phe Gln Leu Tyr Asp Val Arg Asn Val Val Arg Asn 1060
1065 1070 Gly Arg Phe Leu Ser Gly Val Ser Asp
Trp Ile Val Thr Ser Asp Val 1075 1080
1085 Lys Val Gln Glu Glu Asn Gly Asn Asn Val Leu Val Leu Ser
Asn Trp 1090 1095 1100
Asp Ala Gln Val Leu Gln Cys Leu Lys Leu Tyr Gln Asp Arg Gly Tyr1105
1110 1115 1120 Ile Leu Arg Val Thr
Ala Arg Lys Glu Gly Leu Gly Glu Gly Tyr Ile 1125
1130 1135 Thr Ile Thr Asp Glu Glu Gly His Thr Asp
Gln Leu Thr Phe Gly Thr 1140 1145
1150 Cys Glu Glu Ile Asp Ala Ser Asn Thr Phe Val Ser Thr Gly Tyr
Ile 1155 1160 1165 Thr
Lys Glu Leu Glu Phe Phe Pro Asp Thr Glu Lys Val Arg Ile Glu 1170
1175 1180 Ile Gly Glu Thr Glu Gly
Ile Phe Lys Val Glu Ser Val Glu Leu Phe1185 1190
1195 1200 Leu Met Glu Asp Leu Cys 1205
54703PRTBacillus thuringiensis 54Met Arg Leu Lys Lys Leu Leu Val Cys
Asn Ile Arg Ile Gly Gly Thr1 5 10
15 Asn Met Asn Leu Gly Asn Tyr Asn Glu Phe Asp Ile Ile Asp
Ile Thr 20 25 30
Glu Asn Asn Gln Thr Lys Thr Ser Arg Tyr Asn Asn Val Asn Arg Gln 35
40 45 Glu Asn Pro Ser Asn
Met Ile Ile Ser Asn Pro Ser Ser Asn Tyr Pro 50 55
60 Leu Ala Asn Asn Pro Asn Thr Pro Phe Gln
Asn Ile Asn Tyr Lys Asp65 70 75
80 Phe Leu Asn Met Asn Glu Glu Ile Ala Pro Tyr Ala Ser Ser Lys
Asp 85 90 95 Val
Ile Phe Ser Ser Met Asn Ile Ile Arg Thr Phe Met Gly Phe Ala
100 105 110 Gly His Gly Thr Ala
Gly Gly Ile Val Ala Leu Phe Thr Glu Val Leu 115
120 125 Arg Leu Leu Trp Pro Asn Lys Gln Asp
Glu Leu Trp Glu Ser Phe Met 130 135
140 Lys Glu Val Glu Lys Leu Ile Glu Gln Glu Ile Thr Asp
Ala Val Val145 150 155
160 Ser Lys Ala Leu Ala Glu Leu Glu Gly Leu Arg Asn Ala Leu Gln Gly
165 170 175 Tyr Thr Asp Ala
Leu Glu Ala Trp Gln Asn Asn Arg Ser Asp Lys Leu 180
185 190 Lys Gln Leu Leu Val Tyr Asp Arg Phe
Val Ser Thr Glu Asn Leu Phe 195 200
205 Lys Phe Ala Met Pro Ser Phe Arg Val Gly Gly Phe Glu Val
Pro Leu 210 215 220
Leu Thr Val Tyr Ala Gln Ala Ala Asn Leu His Leu Leu Leu Leu Lys225
230 235 240 Asn Ser Glu Leu Phe
Gly Ala Glu Trp Gly Met Gln Gln Tyr Glu Ile 245
250 255 Asp Leu Phe Tyr Asn Glu Gln Lys Asp Tyr
Val Val Glu Tyr Thr Asp 260 265
270 His Cys Val Lys Trp Tyr Thr Glu Gly Leu Asn Arg Leu Lys Asn
Ala 275 280 285 Ser
Gly Val Lys Gly Lys Val Trp Glu Glu Tyr Asn Arg Phe Arg Arg 290
295 300 Glu Met Thr Ile Met Val
Leu Asp Leu Leu Pro Leu Phe Pro Ile Tyr305 310
315 320 Asp Val Arg Thr Tyr Pro Thr Glu Thr Val Thr
Glu Leu Thr Arg Gln 325 330
335 Ile Phe Thr Asp Pro Ile Gly Leu Arg Gly Ile Asn Glu Ser Lys Tyr
340 345 350 Pro Asp Trp
Tyr Gly Ala Ala Ser Asp Ser Phe Ser Leu Ile Glu Asn 355
360 365 Arg Ala Val Pro Gln Pro Ser Leu
Phe Gln Trp Leu Thr Glu Phe Lys 370 375
380 Val Tyr Thr Lys Tyr Val Glu Pro Asn Asp Lys Leu Thr
Ile Leu Ala385 390 395
400 Gly His Ser Val Thr Thr Gln Tyr Thr Ser Tyr Tyr Lys Lys Ser Thr
405 410 415 Phe Thr Tyr Gly
Asp Thr Ser Ser Ala Asn Ser Ser Arg Thr Phe Asp 420
425 430 Leu Leu Ala Lys Asp Val Tyr Gln Val
Asp Ser Val Ala Ala Ala Ser 435 440
445 Lys Ser Ala Thr Trp Tyr Leu Ala Val Pro Glu Met Arg Leu
Tyr Ser 450 455 460
Ile Asn Thr Asn Asn Ile Leu Ser Glu Asp Tyr Phe Ser Leu Ser Thr465
470 475 480 Asn Ile Pro Ser Ser
Lys Met Arg Arg Met Tyr Ser Ser Glu Glu Leu 485
490 495 Pro Ile Gly Ile Ser Asp Thr Pro Ile Tyr
Gly Asp Leu Glu Glu Tyr 500 505
510 Ser His Arg Leu Ser Phe Ile Ser Glu Ile Met His Asn Ser Gly
Ser 515 520 525 Val
Thr Gly Ser Asn Asn Ile Lys Gly Ile Ile Pro Val Leu Gly Trp 530
535 540 Thr His Thr Ser Val Ser
Pro Glu Asn Tyr Ile His Arg Asp Lys Ile545 550
555 560 Ser Gln Leu Tyr Ala Val Lys Ala Tyr Thr Thr
Ser Asn Val Ser Val 565 570
575 Val Gly Gly Pro Gly Phe Leu Gly Gly Asn Ile Ile Lys Gly Asn Asn
580 585 590 Asp Pro Ala
Ser Tyr Thr Gly Ser Val Ser Trp Ala Ile Arg Leu Asp 595
600 605 Gly Ser Thr Val Ser Arg Phe Arg
Leu Arg Ile Pro Tyr Ala Ala Glu 610 615
620 Thr Asp Gly Thr Phe Ser Ile Thr Val Arg Asp Asp Leu
Gly Pro Phe625 630 635
640 Thr Ile Lys Lys Asp Phe Ile Ala Thr Met Lys Pro Gly Asp Pro Leu
645 650 655 Ser Tyr Gly Lys
Phe Glu Tyr Leu Glu Phe Glu Gln Thr Met Ser Leu 660
665 670 Asn Asn Lys His Gly Gln Phe Phe Val
His Thr Glu Asn Leu Lys Asp 675 680
685 Arg Asn Ser Ser Val Tyr Trp Asn Arg Val Glu Ile Ile Pro
Val 690 695 700
55772PRTBacillus thuringiensis 55Met Val Asn Glu Asn Met Asp Met Tyr Asn
Asn Asn Gly Ser Met Asn1 5 10
15 Gly Asn Pro Asp Met Tyr Asn Lys Asn Gly Ser Met Asn Gly Asn
Thr 20 25 30 Asp
Val Tyr Asn Asn Asn Gly Ser Met Asn Gly Asn Pro Asp Met Tyr 35
40 45 Asn Asn Asn Gly Ser Met
Asn Gly Asn Thr Asp Val Tyr Asn Asn Asn 50 55
60 Gly Ser Met Asn Gly Asn Pro Asp Val Tyr Asn
Lys Asn Gly Ser Met65 70 75
80 Asp Gly Asn Pro Asp Met Tyr Asn Asn Asn Gly Ser Met Asn Gly Asn
85 90 95 Thr Asp Val
Tyr Asn Lys Asn Gly Ser Met Asn Gly Asn Pro Asp Met 100
105 110 Tyr Asn Asn Asn Gly Ser Met Asn
Gly Asn Thr Asp Val Tyr Asn Asn 115 120
125 Asn Gly Ser Met Asn Gly Asn Thr Asp Asn Gln Val Pro
Ala Tyr Asn 130 135 140
Ile Leu Ser Ala Glu Asn Pro Ser Asn Ile Leu Glu Ser Asp Thr Arg145
150 155 160 Cys Thr Leu Asn Val
Lys Asn Val Gln Asp Glu Ala Ile Cys Thr Gly 165
170 175 Ser Asn Leu Thr Asn Glu Ile Gly Pro Leu
Val Val Pro Ile Ala Phe 180 185
190 Thr Pro Ile Ile Leu Thr Pro Ala Leu Ile Glu Val Gly Lys Trp
Leu 195 200 205 Gly
Val Gln Ile Gly Lys Trp Ala Leu Ser Thr Ala Leu Lys Glu Leu 210
215 220 Lys Ser Phe Leu Phe Pro
Asn Ser Asp Pro Gln Arg Glu Met Glu Lys225 230
235 240 Leu Arg Ile Glu Leu Glu Asn Ser Phe Asn Lys
Lys Leu Thr Glu Asp 245 250
255 Lys Leu Asn Phe Leu Thr Ala Ala Tyr Thr Gly Phe Asn Asn Leu Ser
260 265 270 Asn Ser Phe
Ile Ser Ala Thr Glu Arg Val Lys Ala Ala Glu Ile Thr 275
280 285 Leu Ala Thr Ala Pro Ser Gln Glu
Asn Gln Asp Ile Leu Asp Glu Ala 290 295
300 Arg Thr Leu Ala Arg Asp Tyr Phe Val Ser Leu His Ser
Gln Met Ile305 310 315
320 Val Trp Leu Pro Gln Phe Glu Ile Ser Gly Tyr Glu Glu Ile Ser Leu
325 330 335 Pro Leu Phe Thr
Gln Met Cys Thr Leu His Leu Thr His Leu Lys Asp 340
345 350 Gly Val Leu Met Gly Gln Asn Trp Gly
Leu Ser Thr Asp Asp Ile Lys 355 360
365 His Phe Lys Gly Glu Phe Tyr Arg Leu Ser Asn Asp Tyr Thr
Ser Arg 370 375 380
Ala Phe Asp Ser Phe His Arg Gly Phe Asn Arg Leu Arg Thr Gln Gln385
390 395 400 Gly Thr Ala Gly Val
Ile Lys Phe Arg Thr Ala Met Asn Ala Tyr Ala 405
410 415 Phe Asp Asn Ile Tyr Lys Trp Ser Leu Leu
Arg Tyr Glu Gly Ile Asn 420 425
430 Pro Arg Ile Thr Arg Ser Leu Trp His Tyr Ile Gly Tyr Asn Ser
Ser 435 440 445 Leu
Gly Ser Asn Asp Phe Asn Thr Leu Tyr Lys Leu Met Val Gly Ile 450
455 460 Pro His Glu Arg Phe Arg
Thr Val Ala Ile Gly Tyr Arg Ala Lys Thr465 470
475 480 Gly Glu Asp Trp Lys Val Thr Gly Ala Lys Ser
Thr Phe Tyr Ser Gly 485 490
495 Gly Gly Glu Trp Val Gly Asn Val Ser Lys Ala Thr Arg Ile Pro Val
500 505 510 Tyr Thr Thr
Lys Thr Asp Trp Arg Gln Phe Glu Arg Arg Ile His Gly 515
520 525 Arg Leu Gly Thr Glu Gln Tyr Thr
Arg Trp His Leu Thr Ile Gln Asp 530 535
540 Thr Asn Ile Ile Gly Asn Ser Tyr Leu Thr Gly Leu Pro
Phe Asp Ile545 550 555
560 Ser Tyr Pro Asp Tyr Phe Ile Arg Thr Ile Ser Ala Lys Pro Glu Ala
565 570 575 Tyr Pro Ile Tyr
Lys Ser Leu Ser Leu Gly Asp Asn Pro Gly Tyr Val 580
585 590 Val Asp Asn Pro Gly Asn Asn Leu Ile
Ile Gly Phe Ser Pro Asp Asn 595 600
605 Leu Lys Thr Phe Met Thr Asp Gly Asn Arg Tyr His Ser Ile
Glu Ser 610 615 620
Gly Tyr Pro Thr Asn Pro Ser Cys Thr Ile Pro Ala Val Leu Tyr Asn625
630 635 640 Ser Val Ser Asn Pro
Phe Gln Ala Tyr Phe Asn Asp Glu Leu Gly Asn 645
650 655 Gly Ser Asp Gly Ser Ile Thr Leu Ile Arg
Arg Gly Gly Ala His Tyr 660 665
670 Leu Val Asp Ser Arg Ser Ala Ser Tyr Asp Arg Ser Phe Arg Leu
Ile 675 680 685 Ile
Arg Ile Gln Ala Gly Ser Ser Ala Phe Lys Val Thr Val Arg Ser 690
695 700 Arg His Thr Ser Glu Ser
Phe Glu Leu Asn Phe Thr Leu Leu Ser Asp705 710
715 720 Gln Asp Ile Asn Tyr Tyr Tyr Asp Tyr Ile Ser
Gln Pro Phe Asn Leu 725 730
735 Ser Ser Thr Tyr Tyr Tyr Ile Asp Val Glu Arg Val Val Ser Asp Asp
740 745 750 Ile Arg Ala
Leu Thr Phe Asn Gln Met Ile Ile Val Pro Thr Thr Glu 755
760 765 Phe Gln Ile Leu 770
56289PRTBacillus thuringiensis 56Met Ala Ile Tyr Asp Ile Ala Ala Asp Leu
Phe Asp Leu Thr Arg Trp1 5 10
15 Tyr Ala Glu Gln Asn Tyr Asn Ala Asn Pro Thr Thr Phe Arg Gly
Ala 20 25 30 Lys
Val Tyr Asp Arg Ile Val Ser Asp Val Gln Ser Ile Pro Glu Lys 35
40 45 Val Asp Phe Asn Leu Ile
Pro Gly Leu Ala Tyr Thr Val Lys Asn Glu 50 55
60 Ile Val Asn Asp Thr Asn Thr Glu Gln Ser Met
Ser Thr Lys Leu Met65 70 75
80 His Thr Leu Ile Glu Ser Asn Ser Val Thr Thr Thr Lys Gly Tyr Lys
85 90 95 Ile Gly Ser
Ser Ile Lys Asn Thr Phe Ser Val Asn Ile Glu Gly Ser 100
105 110 Phe Phe Val Gly Gly Gly Ser Thr
Glu His Ser Ile Glu Val Ser Val 115 120
125 Ser Gly Glu Tyr Asn His Ser Ser Ser Glu Thr Lys Thr
Asn Thr Ser 130 135 140
Gln Lys Thr Trp Glu Tyr Asn Ser Pro Ile Leu Val Pro Ala Lys Thr145
150 155 160 Lys Val Thr Ala Thr
Leu Asp Ile Tyr Ala Gly Pro Val Val Val Pro 165
170 175 Val Thr Leu Lys Ser Thr Val Thr Gly Thr
Gly Ile Val Asn Asn Phe 180 185
190 Pro Asn Val Leu Thr Ser Leu Ser Tyr Ile Asp Arg Asn Asn Lys
Leu 195 200 205 Trp
Thr Asp Ser Leu Pro Thr Ala Leu Leu Tyr Asp Tyr Arg Asn Gln 210
215 220 Trp Pro Gly Ser Gln Ser
Ile Tyr Val Gly Lys Asn Gly Gly Gly Val225 230
235 240 Gln Val Glu Gly Lys Ala Glu Ile Gln Leu Glu
Leu Gly Leu Tyr Ser 245 250
255 Ile Ala Thr Phe Asp Ser Gln Pro Leu Ser Gly Asn Thr Thr Gly Lys
260 265 270 Glu Ala Val
Tyr Ser Lys Ala Ile Leu Arg Asp Gly Ser Ile Ile Asp 275
280 285 Ile57295PRTBacillus
thuringiensis 57Met Ala Ile His Asp Val Gly Thr Asp Leu Leu Glu Phe Ala
Lys Trp1 5 10 15
Tyr Ala Thr Thr Asn Tyr Asn Ala Asn Pro Asn Thr Phe Arg Asn Pro
20 25 30 Gln Ile Phe Glu Ser
Val Val Gly Glu Ser Glu Ile Ile Pro Lys Asn 35 40
45 Asp Thr Phe Glu Thr Ile Pro Lys Leu Thr
Thr Val Val Thr Asp Val 50 55 60
Ile Ile Asn Asp Thr Ser Val Pro Gln Ser Ile Thr Pro Lys Ile
Met65 70 75 80 Gln
Lys Thr Ser Glu Thr Ile Thr Thr Thr Thr Thr Gln Gly Phe Lys
85 90 95 Val Gly Ser Glu Ile Lys
Tyr Thr Asn Thr Met Lys Val Asn Leu Leu 100
105 110 Leu Val Gly Gly Val Ser Asn Ser Ile Ala
Val Ser Ile Ser Ala Glu 115 120
125 Tyr Asn Tyr Ser Ser Ser Glu Thr Glu Thr Asn Ile Thr Glu
Lys Ala 130 135 140
Trp Glu Tyr Asn Arg Pro Val Leu Val Leu Pro Arg Thr Lys Val Thr145
150 155 160 Ala Thr Leu Ser Ile
Tyr Ser Gly Ser Phe Thr Ile Pro Val Thr Leu 165
170 175 Lys Ser Thr Ile Ser Gly Asn His Ile Ser
Asn Ser Gly Tyr Gly Tyr 180 185
190 Ala Leu Ser Ser Ile Gly Tyr Thr Asp Tyr Asn Asn Arg Ser Trp
Thr 195 200 205 Asp
Ile Tyr Arg Thr Asn Phe Leu Tyr Asp Tyr Arg Asn Glu Trp Pro 210
215 220 Gly Arg Lys Pro Ile Tyr
Val Gly Arg Asp Asn Ile Gly Val Lys Val225 230
235 240 Glu Gly Glu Ser Arg Val Asp Ala Glu Leu Gly
Leu Tyr Ser Ile Val 245 250
255 Thr Phe Lys Glu Glu Pro Leu Pro Gly Asn Asn Leu Ile Gly Asn Gly
260 265 270 Arg Thr Tyr
Ser Met Ala Ile Leu Arg Asp Gly Ser Thr Met Asp Ile 275
280 285 Ser Ile Pro Lys Asn Asn Asn
290 295 581333PRTBacillus thuringiensis 58Met Ala Gln Leu
Asn Glu Ile Tyr Pro Ser Tyr Tyr Asn Val Leu Ala1 5
10 15 Tyr Pro Pro Leu Ile Leu Asp Asp Lys
Ser Leu Tyr Asp Gln Tyr Thr 20 25
30 Glu Trp Lys Lys Lys Ile Asp Lys Thr Trp Lys Gln Tyr Asp
Lys Asp 35 40 45
Phe Leu Pro Lys Pro Leu Met Asp Leu Gly Lys Ser Leu Ala Glu Ala 50
55 60 Tyr Lys Gly Asp Pro
Asp Gly Tyr Leu His Ile Ala Asn Thr Ala Ile65 70
75 80 Arg Ile Ala Phe Leu Leu Ile Pro Gly Gly
Gln Thr Ala Ala Phe Gly 85 90
95 Val Asn Leu Val Leu Asn Lys Ala Ile Gly Ile Phe Tyr Pro Pro
Gln 100 105 110 Asn
Lys Ser Leu Phe Asp Gln Ile Lys Asp Ala Val Ser Asn Leu Val 115
120 125 Asp Gln Lys Leu Ile Asp
Gln Glu Ile Ser Gly Val Leu Ile Lys Leu 130 135
140 Asn Ser Leu Gln Gln Pro Leu Ser Arg Phe Ser
Asn Ser Ile Gln Arg145 150 155
160 Ala Val Gly Lys Pro Gln Asp Phe Asp Asp Gln Thr Thr Ser Ser Asn
165 170 175 Ala Ile Ile
Leu Asp Glu Thr Gln Asp Cys Ser Lys Asp Asp Ser Cys 180
185 190 Ser Cys Ser Asn Thr Gln Pro Arg
Pro Ser Asp Ala Pro Leu Cys Thr 195 200
205 Pro Cys Ile Cys Arg Met Lys Glu Val Gln Gln Thr Phe
Asn Asn Ser 210 215 220
Ser Thr Asp Val Asn Arg Ala Leu Thr Asp Met Lys Thr Thr Leu Lys225
230 235 240 Asp Val Val Gly Ala
Asp Gln Leu Arg Ser Tyr Met Gln Ile Tyr Leu 245
250 255 Pro Leu Tyr Val Thr Ala Ala Thr Met Glu
Leu Gln Met Tyr Lys Thr 260 265
270 Tyr Ile Asp Phe Thr Gln Lys Phe Asp Phe Asp Val Thr Gly Thr
Thr 275 280 285 Lys
Glu His Val Asn Glu Leu Arg Gln Lys Ile Lys Thr His Ser Glu 290
295 300 Tyr Ile Met Gly Leu Phe
Lys Lys Ser Leu Pro Glu Ile Ser Asn Asn305 310
315 320 Thr Lys Glu Gln Leu Asn Ala Tyr Ile Lys Tyr
Thr Arg Asn Ile Thr 325 330
335 Leu Asn Ala Leu Asp Met Val Ser Thr Trp Lys Phe Leu Asp Pro Val
340 345 350 Asp Tyr Pro
Thr Thr Ala Thr Phe Asn Pro Thr Arg Ile Ile Phe Asn 355
360 365 Asp Leu Ala Gly Pro Val Glu Cys
Leu Asn Ser Thr Gln Asp Ser Asn 370 375
380 Lys Leu His Phe Asn Phe Phe Asp Met Asn Gly Gln Ser
Met Pro Asn385 390 395
400 Asn Asp Ile Phe Asn Tyr Phe Tyr Arg Gly Met Gln Val Lys Gly Leu
405 410 415 Gln Ile Gln Thr
Tyr Thr Ser Ser Asp Thr Lys Asn Pro Gln His Phe 420
425 430 Pro Val Gly Phe Leu Ser Ser Tyr Tyr
Gly Ser Asn Gly Asp Phe Pro 435 440
445 Phe Asp Lys Arg Val Asp Pro Asn Lys Phe Thr Gly Gly Ser
Lys Ser 450 455 460
Val Lys Leu Gly Asp Asp Val Tyr Glu Ser Arg Ser Ala Leu Ser Val465
470 475 480 Ile Asn Ala Val Ser
Asn Gln Leu Gln Val Phe Leu Asn Tyr Ile Asp 485
490 495 Thr Glu Asp Leu Tyr Phe Asp Gln Ser Val
Ser Pro Gly Gly Thr Ala 500 505
510 Cys Gly Ser Gly Asn Ser Thr Ile Trp Pro Asp Gln Lys Ile Gln
Ala 515 520 525 Ile
Tyr Pro Ile Gln Pro Asp Asn Ser Gln Thr Tyr Pro Ser Tyr Tyr 530
535 540 Ser Thr Ser Lys Ile Gly
Phe Val Thr Thr Leu Val Pro Asn Asp Thr545 550
555 560 Thr Pro Trp Ile Thr Phe Thr Asp Asn Gly Asn
Asn Ser Ile Tyr Thr 565 570
575 Phe Ser Ala Glu Asn Thr Arg Thr Leu Thr Gly Ser Ala Gly Pro Val
580 585 590 Arg Glu Phe
Ile Thr Gly Ser Ala Pro Leu Gly Leu Ser Pro Gly Gly 595
600 605 Gly Ala Gln Tyr Ser Ile Asn Thr
Ser Asp Ala Pro Ser Gly Asp Tyr 610 615
620 Gln Val Arg Val His Val Ala Thr Pro Gly Ser Gly Gly
Ser Leu Ala625 630 635
640 Ile Ser Val Asp Gly Lys Thr Gln Thr Leu Gln Leu Pro Asp Thr Asn
645 650 655 Val Asn Asp Thr
Asn His Ile Ala Gly Phe Ala Gly Thr Tyr Thr Leu 660
665 670 Ala Pro Ala Thr Gln Val Asp Ala Ala
Thr Leu Lys Pro Lys Ala Pro 675 680
685 Thr Glu Asn Ile Phe Pro Val Arg Gln Thr Ser Ser Leu Pro
Val Ser 690 695 700
Ile Thr Asn Asn Ser Ser Thr Val Ile Asn Ile Asp Arg Ile Glu Phe705
710 715 720 Val Pro Val Ser Ala
Pro Ala Pro Asp Pro Ser Pro Asp Ser Gly Lys 725
730 735 Pro Ile His Lys Ser Val Pro Lys Thr Val
Thr Gln Leu Ser Thr Thr 740 745
750 Lys Glu Ile Trp Ser Ser Thr Ser Glu Tyr Ala Thr Thr Ile Ser
Phe 755 760 765 Thr
Gly Asn Val Tyr Asn Asp Ala Ser Ile Thr Phe Gln Leu Leu Ser 770
775 780 Ser Gly Gln Val Val Lys
Glu Phe Pro Phe Thr Gly Asn Gly Val Ala785 790
795 800 Ser Lys Pro Gly Phe His Gly Ser Ser Pro Ser
Cys Tyr Asp Thr Pro 805 810
815 Tyr Pro Phe Ser Gln Pro Asp Leu Ser Val Pro Lys Tyr Asn Lys Leu
820 825 830 Gln Val Val
Met Lys Ser Asp Gly Tyr Ser Lys Pro Cys Asp Leu Gly 835
840 845 Asp Ser Phe Pro Asn Thr Phe Asp
Ala Glu Ile Asp Ile Lys Phe Asn 850 855
860 Leu Ser Asp Thr Ala Asp Leu Ala Gln Ile Thr Ala Gln
Val Gln Gly865 870 875
880 Leu Phe Thr Ser Ser Ser Ser Thr Glu Leu Ser Pro Asn Val Ser Gly
885 890 895 Tyr Gln Ile Asp
Gln Ile Ala Leu Lys Val Asn Ala Leu Ser Asp Glu 900
905 910 Val Phe Cys Lys Glu Lys Ile Val Leu
Arg Lys Leu Val Asn Lys Ala 915 920
925 Lys Gln Phe Met Lys Thr Arg Asn Leu Leu Ile Gly Gly Asp
Phe Glu 930 935 940
Ile Leu Asp Lys Trp Ala Leu Gly Thr Gln Ala Thr Ile Lys Asp Asn945
950 955 960 Ser Ser Leu Phe Lys
Gly Asn His Leu Phe Leu Gln Pro Thr Asn Gly 965
970 975 Ile Ser Ser Ser Tyr Ala Tyr Gln Lys Ile
Asp Glu Ser Lys Leu Lys 980 985
990 Pro Tyr Thr Arg Tyr Asn Val Ser Gly Phe Val Ala Gln Ser Glu
His 995 1000 1005 Leu
Glu Ile Val Val Ser Arg Tyr Gly Lys Glu Ile Asp Lys Ile Leu 1010
1015 1020 Asn Val Pro Tyr Glu Glu
Ala Leu Pro Val Ser Ser Gly Asn Gln Ser1025 1030
1035 1040 Thr Cys Cys Lys Pro Ser Ser Cys Ser Cys Ser
Ala Cys Thr Gly Gly 1045 1050
1055 Pro His Pro His Phe Phe Ser Tyr Ser Ile Asp Val Gly Lys Leu Tyr
1060 1065 1070 Pro Asp Leu
Asn Pro Gly Ile Glu Phe Gly Leu Arg Leu Ala His Pro 1075
1080 1085 Ser Gly Tyr Ala Lys Val Gly Asn
Leu Glu Ile Val Glu Glu Arg Pro 1090 1095
1100 Leu Thr Asn Thr Glu Ile Arg Lys Ile Gln Arg Lys Glu
Glu Lys Trp1105 1110 1115
1120 Lys Lys Ala Trp Asp Thr Glu Arg Ala Glu Ile Asn Ala Ile Leu Gln
1125 1130 1135 Pro Val Ile Asn
Gln Ile Asn Ala Phe Tyr Thr Asn Gly Asp Trp Asn 1140
1145 1150 Gly Ser Ile Leu Pro His Val Thr Tyr
Gln Asp Leu Tyr Asn Ile Val 1155 1160
1165 Leu Pro Glu Leu Ser Lys Leu Arg His Trp Phe Met Lys Asp
Arg Pro 1170 1175 1180
Gly Glu His Tyr Thr Ile Leu Gln Gln Phe Lys Gln Ala Leu Glu Arg1185
1190 1195 1200 Val Phe Asn Gln Leu
Glu Glu Arg Asn Leu Ile His Asn Gly Ser Phe 1205
1210 1215 Thr Asn Gly Leu Ala Asn Trp Leu Val Asp
Gly Asp Thr Gln Ile Thr 1220 1225
1230 Thr Leu Glu Asn Gly Asn Leu Ala Leu Gln Leu Ser Asp Trp Asp
Ala 1235 1240 1245 Ser
Ala Ser Gln Ser Ile Asp Ile Ser Asp Phe Asp Glu Asp Lys Glu 1250
1255 1260 Tyr Thr Val Arg Val Tyr
Ala Lys Gly Lys Gly Thr Ile Arg Thr Val1265 1270
1275 1280 Asn Cys Glu Asn Glu Pro Leu Ser Phe Asn Thr
Asn Thr Phe Thr Ile 1285 1290
1295 Leu Glu Gln Arg Leu Tyr Phe Asp Asn Pro Ser Val Leu Leu His Ile
1300 1305 1310 Gln Ser Glu
Gly Ser Glu Phe Val Ile Gly Ser Val Glu Leu Ile Glu 1315
1320 1325 Leu Ser Asp Asp Glu 1330
59723PRTBacillus thuringiensis 59Met Ala Gln Leu Asn Glu Ile Tyr
Pro Ser Tyr Tyr Asn Val Leu Ala1 5 10
15 Tyr Pro Pro Leu Ile Leu Asp Asp Lys Ser Leu Tyr Asp
Gln Tyr Thr 20 25 30
Glu Trp Lys Lys Lys Ile Asp Lys Thr Trp Lys Gln Tyr Asp Lys Asp
35 40 45 Phe Leu Pro Lys
Pro Leu Met Asp Leu Gly Lys Ser Leu Ala Glu Ala 50 55
60 Tyr Lys Gly Asp Pro Asp Gly Tyr Leu
His Ile Ala Asn Thr Ala Ile65 70 75
80 Arg Ile Ala Phe Leu Leu Ile Pro Gly Gly Gln Thr Ala Ala
Phe Gly 85 90 95
Val Asn Leu Val Leu Asn Lys Ala Ile Gly Ile Phe Tyr Pro Pro Gln
100 105 110 Asn Lys Ser Leu Phe
Asp Gln Ile Lys Asp Ala Val Ser Asn Leu Val 115
120 125 Asp Gln Lys Leu Ile Asp Gln Glu Ile
Ser Gly Val Leu Ile Lys Leu 130 135
140 Asn Ser Leu Gln Gln Pro Leu Ser Arg Phe Ser Asn Ser
Ile Gln Arg145 150 155
160 Ala Val Gly Lys Pro Gln Asp Phe Asp Asp Gln Thr Thr Ser Ser Asn
165 170 175 Ala Ile Ile Leu
Asp Glu Thr Gln Asp Cys Ser Lys Asp Asp Ser Cys 180
185 190 Ser Cys Ser Asn Thr Gln Pro Arg Pro
Ser Asp Ala Pro Leu Cys Thr 195 200
205 Pro Cys Ile Cys Arg Met Lys Glu Val Gln Gln Thr Phe Asn
Asn Ser 210 215 220
Ser Thr Asp Val Asn Arg Ala Leu Thr Asp Met Lys Thr Thr Leu Lys225
230 235 240 Asp Val Val Gly Ala
Asp Gln Leu Arg Ser Tyr Met Gln Ile Tyr Leu 245
250 255 Pro Leu Tyr Val Thr Ala Ala Thr Met Glu
Leu Gln Met Tyr Lys Thr 260 265
270 Tyr Ile Asp Phe Thr Gln Lys Phe Asp Phe Asp Val Thr Gly Thr
Thr 275 280 285 Lys
Glu His Val Asn Glu Leu Arg Gln Lys Ile Lys Thr His Ser Glu 290
295 300 Tyr Ile Met Gly Leu Phe
Lys Lys Ser Leu Pro Glu Ile Ser Asn Asn305 310
315 320 Thr Lys Glu Gln Leu Asn Ala Tyr Ile Lys Tyr
Thr Arg Asn Ile Thr 325 330
335 Leu Asn Ala Leu Asp Met Val Ser Thr Trp Lys Phe Leu Asp Pro Val
340 345 350 Asp Tyr Pro
Thr Thr Ala Thr Phe Asn Pro Thr Arg Ile Ile Phe Asn 355
360 365 Asp Leu Ala Gly Pro Val Glu Cys
Leu Asn Ser Thr Gln Asp Ser Asn 370 375
380 Lys Leu His Phe Asn Phe Phe Asp Met Asn Gly Gln Ser
Met Pro Asn385 390 395
400 Asn Asp Ile Phe Asn Tyr Phe Tyr Arg Gly Met Gln Val Lys Gly Leu
405 410 415 Gln Ile Gln Thr
Tyr Thr Ser Ser Asp Thr Lys Asn Pro Gln His Phe 420
425 430 Pro Val Gly Phe Leu Ser Ser Tyr Tyr
Gly Ser Asn Gly Asp Phe Pro 435 440
445 Phe Asp Lys Arg Val Asp Pro Asn Lys Phe Thr Gly Gly Ser
Lys Ser 450 455 460
Val Lys Leu Gly Asp Asp Val Tyr Glu Ser Arg Ser Ala Leu Ser Val465
470 475 480 Ile Asn Ala Val Ser
Asn Gln Leu Gln Val Phe Leu Asn Tyr Ile Asp 485
490 495 Thr Glu Asp Leu Tyr Phe Asp Gln Ser Val
Ser Pro Gly Gly Thr Ala 500 505
510 Cys Gly Ser Gly Asn Ser Thr Ile Trp Pro Asp Gln Lys Ile Gln
Ala 515 520 525 Ile
Tyr Pro Ile Gln Pro Asp Asn Ser Gln Thr Tyr Pro Ser Tyr Tyr 530
535 540 Ser Thr Ser Lys Ile Gly
Phe Val Thr Thr Leu Val Pro Asn Asp Thr545 550
555 560 Thr Pro Trp Ile Thr Phe Thr Asp Asn Gly Asn
Asn Ser Ile Tyr Thr 565 570
575 Phe Ser Ala Glu Asn Thr Arg Thr Leu Thr Gly Ser Ala Gly Pro Val
580 585 590 Arg Glu Phe
Ile Thr Gly Ser Ala Pro Leu Gly Leu Ser Pro Gly Gly 595
600 605 Gly Ala Gln Tyr Ser Ile Asn Thr
Ser Asp Ala Pro Ser Gly Asp Tyr 610 615
620 Gln Val Arg Val His Val Ala Thr Pro Gly Ser Gly Gly
Ser Leu Ala625 630 635
640 Ile Ser Val Asp Gly Lys Thr Gln Thr Leu Gln Leu Pro Asp Thr Asn
645 650 655 Val Asn Asp Thr
Asn His Ile Ala Gly Phe Ala Gly Thr Tyr Thr Leu 660
665 670 Ala Pro Ala Thr Gln Val Asp Ala Ala
Thr Leu Lys Pro Lys Ala Pro 675 680
685 Thr Glu Asn Ile Phe Pro Val Arg Gln Thr Ser Ser Leu Pro
Val Ser 690 695 700
Ile Thr Asn Asn Ser Ser Thr Val Ile Asn Ile Asp Arg Ile Glu Phe705
710 715 720 Val Pro
Val60378PRTBacillus thuringiensis 60Met Tyr Cys Asn Thr Ile Leu Arg Lys
Arg Tyr Lys Lys Leu Ala Thr1 5 10
15 Ile Ile Pro Leu Thr Ser Met Ser Ala Val Ala Ile Ala Pro
Ala Thr 20 25 30
Ser Phe Ala Val Glu Thr Gln Lys Ala Asp Val Ser Ser Gln Glu Gly 35
40 45 Pro Ile Gln Gly Tyr
Gln Met Glu Asn Gly Lys Ile Thr Pro Val Tyr 50 55
60 Lys Asn Lys Leu Thr Gln Phe Asn Thr Ala
Asp Asp Ile Asp Pro Gly65 70 75
80 Leu Pro Leu Leu Pro Glu Asn Pro Tyr Asn Pro Ile Pro Asp His
Gly 85 90 95 Thr
Ala Tyr Val Glu Ser Thr Asp Ile Gly Asp Thr Val Tyr Phe Lys
100 105 110 Pro Phe Glu Pro Pro
Lys Asn Asn Val Leu Glu Leu Gly Asp Cys Asp 115
120 125 Asp Asn Thr Tyr Gln Trp Ser Val Phe
Val Asp Ser Gln Lys Tyr Lys 130 135
140 Ser Val Gly Tyr Phe Val Gln Lys Gln Ala Asp Gly Gln
Ile Arg Val145 150 155
160 Gly Tyr Tyr Asn Pro Glu Asp Leu Ser Leu Ile Thr Asp Ser Asn His
165 170 175 Ala Phe Ala Gly
Val Pro Gly Phe Lys Leu Thr Ala Glu Glu Lys Ala 180
185 190 Glu Met Gln Arg Asp Leu Asn Arg Glu
Tyr Gly Asp Ile Trp Asp Gly 195 200
205 Thr Ser Lys Leu Lys Arg Glu Thr Asn Tyr Lys Leu Leu Pro
Asn Ala 210 215 220
Ser Gly Leu Gln Asp Asp Ala Ser Gly Phe Gly Tyr Asn Gln Thr Leu225
230 235 240 Thr Ser Gly Val Ser
Thr Thr Asn Met Phe Gly Ile Ala Thr Thr Val 245
250 255 Gly Trp Lys Met Gly Ile Lys Val Ser Val
Val Pro Leu Val Ala Asp 260 265
270 Val Thr Ser Glu Ile Ser Ala Ser Leu Thr Ala Ser Tyr Gln His
Thr 275 280 285 Val
Asn Val Thr Asn Gln Thr Ser Ser Gln Val Lys Phe Asp Val Ser 290
295 300 Arg Val Asp Asn Pro Asp
Tyr Lys Tyr Asn Asp Tyr Ala Ala Ala Val305 310
315 320 Tyr Lys Ile Tyr Thr Asp Tyr Thr Leu Glu Pro
Gly Lys Gly Leu Ser 325 330
335 Arg Phe Leu Ala Lys Gln Asp Leu Lys Asp Pro Val Arg Thr Ala Ala
340 345 350 Leu Ala Asn
Thr Asn Tyr Ala Tyr Glu Gly Ser Lys Tyr Tyr Phe Thr 355
360 365 Val Thr Pro Gly Ser His Lys Lys
Ile Val 370 375 61383PRTBacillus
thuringiensis 61Met Asp Phe Leu Asn Tyr Tyr Asn Lys Leu Lys Asn Glu Leu
Asp Asp1 5 10 15
Val Asn Ser Lys Lys Tyr Ser Leu Glu Tyr Thr Ser Asp Gly Leu Met
20 25 30 Val Gln Pro Thr Asp
Asp Pro Leu Asn Thr Met Pro Leu Pro Asp Arg 35 40
45 Pro Val Leu Ser Gly Asn Pro Asn Asp Pro
Ile Pro Ser Glu Gly Thr 50 55 60
Thr Arg Thr Asp Ile Gln Lys Gln Asn Pro Pro Phe Phe Thr Phe
Lys65 70 75 80 Val
Val Ala Lys Leu Ala Tyr Ser Gly Lys Gly Glu Asn Cys Gln Lys
85 90 95 Ala Arg Ala Ala Ser Val
Tyr Gly Ala Val Leu Glu Leu Glu Lys Val 100
105 110 Lys Gln Leu Pro Glu Tyr Ser Asn Val Tyr
Leu Tyr Ser Glu Thr Gly 115 120
125 Ile Lys Thr Asp Arg Ser Asn Ile Arg Tyr Asn Thr Asp Gly
Ile Ile 130 135 140
Gln Phe Leu Asn Pro Ser Phe Ile Asn Thr Phe Ser Ser Asn Pro Ile145
150 155 160 Lys Tyr Gly Asp Thr
Val Gly Tyr Ile Ser Tyr Pro Tyr Asp Thr Leu 165
170 175 Lys Phe Pro Ser Thr Thr Gln Leu Glu Arg
Leu Val Tyr Phe Asn Leu 180 185
190 Leu Asp Ser Asn Ile Leu Asp Lys His Ile Gly Phe Asp Trp Ser
Lys 195 200 205 Ser
Val Thr Asn Gly Thr Glu Asp Thr Glu Met Trp Thr His Ser Ser 210
215 220 Thr Val Gly Ala Glu Leu
Asn Leu Lys Asp Ile Leu Gln Ile Asn Ala225 230
235 240 Ser Tyr Glu His Thr Phe Ser Thr Ser His Met
Glu Lys Lys Glu Asn 245 250
255 Thr Val Ser Lys Thr Ala His Phe Asn Ser Pro Leu Pro Pro Tyr Asn
260 265 270 Tyr Ala Thr
Trp Val Ala Ala Ile Tyr Gln Leu Ser Ile Arg Tyr Gln 275
280 285 Arg Thr Asn Ala Gln Pro Ile Leu
Asp Thr Ile Asn Ala Val Asn Ser 290 295
300 Gly Leu Thr Ala Ser Glu Thr Asp Ile Tyr Leu Lys Ala
Leu Tyr Gly305 310 315
320 Ala Gly Lys Asn Gly Lys Pro Ala Val Gly Asp Pro Ser Ile Leu His
325 330 335 Lys Leu Ser Asn
Val Ile Glu Asp Ala Tyr Glu Tyr Leu Tyr Tyr Ser 340
345 350 Asp Thr Leu Tyr Phe Thr Gln Thr Pro
Ser Gly Asn Ser Pro Thr Pro 355 360
365 Asn Ser Pro Asn Arg Ile Gln Phe Ile Ala Thr Asp Pro Gln
Ser 370 375 380
62634PRTArtificial Sequencetruncated variant Axmi207 62Met Pro Ser Ser
Asp Ser Phe Leu Tyr Ser His Asn Asn Tyr Pro Tyr 1 5
10 15 Ala Thr Asp Pro Asn Thr Val Leu Glu
Gly Arg Asn Tyr Lys Glu Trp 20 25
30 Leu Asn Lys Cys Thr Asp Asn Tyr Thr Asp Ala Leu Gln Ser
Pro Glu 35 40 45
Ala Thr Ala Ile Ser Lys Gly Ala Val Ser Ala Ala Ile Ser Ile Ser 50
55 60 Thr Lys Val Leu Gly
Leu Leu Gly Val Pro Phe Ala Ala Gln Ile Gly 65 70
75 80 Gln Leu Trp Thr Phe Ile Leu Asn Ala Leu
Trp Pro Ser Asp Asn Thr 85 90
95 Gln Trp Glu Glu Phe Met Arg His Val Glu Glu Leu Ile Asn Gln
Arg 100 105 110 Ile
Ala Asp Tyr Ala Arg Asn Lys Ala Leu Ala Glu Leu Thr Gly Leu 115
120 125 Gly Asn Asn Leu Asp Leu
Tyr Ile Glu Ala Leu Asp Asp Trp Lys Arg 130 135
140 Asn Pro Thr Ser Gln Glu Ala Lys Thr Arg Val
Ile Asp Arg Phe Arg 145 150 155
160 Ile Val Asp Gly Leu Phe Glu Ala Tyr Ile Pro Ser Phe Ala Val Ser
165 170 175 Gly Tyr
Gln Val Gln Leu Leu Thr Val Tyr Ala Ala Ala Ala Asn Leu 180
185 190 His Leu Leu Leu Leu Arg Asp
Ser Thr Ile Tyr Gly Ile Asp Trp Gly 195 200
205 Leu Ser Gln Thr Asn Val Asn Asp Asn Tyr Asn Arg
Gln Ile Arg Leu 210 215 220
Thr Ala Thr Tyr Ala Asn His Cys Thr Thr Trp Tyr Gln Thr Gly Leu 225
230 235 240 Glu Arg Leu
Arg Gly Ser Asn Ala Ser Ser Trp Val Thr Tyr Asn Arg 245
250 255 Phe Arg Arg Glu Met Thr Leu Thr
Val Leu Asp Ile Cys Ser Leu Phe 260 265
270 Ser Asn Tyr Asp Tyr Arg Ser Tyr Pro Ala Glu Val Arg
Gly Glu Ile 275 280 285
Thr Arg Glu Ile Tyr Thr Asp Pro Val Gly Val Gly Trp Val Asp Ser 290
295 300 Ala Pro Ser Phe
Gly Glu Ile Glu Asn Leu Ala Ile Arg Ala Pro Arg 305 310
315 320 Thr Val Thr Trp Leu Asn Ser Thr Arg
Ile Phe Thr Gly Arg Leu Gln 325 330
335 Gly Trp Ser Gly Thr Asn Asn Tyr Trp Ala Ala His Met Gln
Asn Phe 340 345 350
Ser Glu Thr Asn Ser Gly Asn Ile Gln Phe Glu Gly Pro Leu Tyr Gly
355 360 365 Ser Thr Val Gly
Thr Ile His Arg Thr Asp Asp Tyr Asp Met Gly Asn 370
375 380 Arg Asp Ile Tyr Thr Ile Thr Ser
Gln Ala Val Leu Gly Leu Trp Ala 385 390
395 400 Thr Gly Gln Arg Val Leu Gly Val Ala Ser Ala Arg
Phe Thr Leu Arg 405 410
415 Asn Leu Phe Asn Asn Leu Thr Gln Val Leu Val Tyr Glu Asn Pro Ile
420 425 430 Ser Ser Thr
Phe Gly Ser Ser Thr Leu Thr His Glu Leu Ser Gly Glu 435
440 445 Asn Ser Asp Arg Pro Thr Ser Ser
Asp Tyr Ser His Arg Leu Thr Ser 450 455
460 Ile Thr Gly Phe Arg Ala Gly Ala Asn Gly Thr Val Pro
Val Phe Gly 465 470 475
480 Trp Thr Ser Ala Thr Val Asp Arg Asn Asn Ile Ile Glu Arg Asn Lys
485 490 495 Ile Thr Gln Phe
Pro Gly Val Lys Ser His Thr Leu Asn Asn Cys Gln 500
505 510 Val Val Arg Gly Thr Gly Phe Thr Gly
Gly Asp Trp Leu Arg Pro Asn 515 520
525 Asn Asn Gly Thr Phe Arg Leu Thr Ile Thr Ser Phe Ser Ser
Gln Ser 530 535 540
Tyr Arg Ile Arg Leu Arg Tyr Ala Thr Ser Val Gly Asn Thr Ser Leu 545
550 555 560 Val Ile Ser Ser Ser
Asp Ala Gly Ile Ser Ser Thr Thr Ile Pro Leu 565
570 575 Thr Ser Thr Ile Thr Ser Leu Pro Gln Thr
Val Pro Tyr Gln Ala Phe 580 585
590 Arg Val Val Asp Leu Pro Ile Thr Phe Thr Thr Pro Thr Thr Gln
Arg 595 600 605 Asn
Tyr Thr Phe Asp Phe Arg Leu Gln Asn Pro Ser Asn Ala Asn Val 610
615 620 Phe Ile Asp Arg Phe Glu
Phe Val Pro Ile 625 630 63306DNABacillus
thuringiensis 63atgacaacaa taaatgaatt atatccggct gtaccttata atgtactggc
atatgctcca 60ccacttaatt tagctgattc gacaccatgg ggtcaaatag ttgttgctga
tgcaattaaa 120gaagcttggg ataattttca aaaatatggt gtattagatt taacagctat
aaatcaaggg 180tttgatgatg caaatacagg ttcttttagt tatcaagctt taatacaaac
tgttttgggt 240attataggta caatttggta tgacagttcc tgtggctgct ccatttgcag
ctacagcgcc 300tattat
306644065DNABacillus thuringiensis 64atgacagttc ctgtggctgc
tccatttgca gctacagcgc ctattattag tttatttgta 60ggattttttt ggcctaaaaa
agataaggga ccacaattaa tcgatataat tgataaagaa 120attaaaaaat tattagataa
ggaattagga gagcaaaaac gtaatgattt agttagtgct 180ttaaatgaga tgcaagaggg
agcaaatgag ttaagtgata ttatgactaa tgcacttttt 240gaaggtacta tacagggaaa
tgttgttact aatgataacc ctcaaggtaa aaggcgaact 300cctaaagctc caacagttag
tgattatgag aatgtttatt cggcatattt tgtggaacat 360gtggatttta gaaacaaaat
atctacgttt cttactggtt cttatgatct tatagcactc 420ccattatatg cattagcaaa
aacaatggag ctttcattgt atcaatcatt tattaatttt 480gctaataaat ggatggattt
tgtatataca aaagcaatta atgaatcagc aactgatgat 540atgaaaagag attatcaagc
gagatacaat actcaaaaaa gtaatttagc tgtacaaaaa 600acacaattga ttaacaaaat
taaagatggt acagatgctg ttatgaaagt ttttaaagat 660accaataatt taccttcaat
aggtactaat aaattagcag taaatgctcg taataagtat 720attagggcct tacaaataaa
ttgtttagat ttagttgctt tgtggcctgg cttatatcca 780gatgaatatc ttttaccatt
acaattagat aaaacacgtg ttgtattttc tgatacaatg 840ggacctgatg aaacacatga
tggtcaaatg aaagttttaa atatattaga ctcaactaca 900agttataacc atcaagatat
aggaataagt acaactcaag atgtaaattc tttattattt 960tatccaagaa aagaactgtt
agaattagat tttgctaaat atatttcatc tagtagtcgt 1020ttttgggttt atggatttgg
cttaaaatat tcagatgata acttttatag atatggtgat 1080aacgatccaa gcagtgattt
taaacctgca tataagtggt ttacgaaaaa ttcccagttc 1140gaaaaccttc ctacttatgg
aaatcctact cctattacta atttaaatgc taaaactcaa 1200gtaacttctt atcttgatgc
attaatatat tatatagacg gaggaactaa tctatataat 1260aatgcgattc ttcatgatac
agggggttat attccgggat atccaggtgt agaaggatat 1320ggtatgagta ataatgaacc
tttagcagga caaaaattaa atgctttata tcctataaaa 1380gtggaaaatg taagtggttc
acaaggaaaa ttaggaacaa tagcagctta tgttccttta 1440aatttacaac cagaaaatat
tattggtgat gctgatccga atacaggttt tccccttaat 1500gtaattaaag gatttccatt
tgaaaaatat ggacctgatt atgagggacg aggaatttcg 1560gttgtaaaag aatggataaa
tggtgcaaat gctgtaaaat tgtctccagg tcaatcagtt 1620ggggtacaaa ttaaaaatat
aacaaaacaa aattatcaaa ttcgtactcg ttatgcaagt 1680aataacagta atcaagtata
ttttaatgta gatccaggtg gatcaccatt atttgcacaa 1740tcagtaacat ttgaatctac
aacaaatgtt acaagtggcc aacaaggcga aaatggtaga 1800tatacattaa aaactatttt
ttctggtaat gatctactta cagtagaaat ccctgttgga 1860aatttttatg tgcatgttac
gaataaagga tcttctgata tctttttaga tcgtcttgag 1920ttttctacag ttccttcata
tgttatatat tcaggtgatt atgatgctac aggtacagat 1980gatgtcttat tgtcagatcc
acatgagtat ttttatgatg tcatagtgaa tggtactgct 2040agtcattcta gtgcagctac
ttctatgaat ttgctcaata aaggaaccgt agtaagaagc 2100attgatattc caggtcactc
aacgtcttat tctgtacagt attcagttcc agaaggattt 2160gatgaagtta gaattctcag
ttctcttccg gatattagtg gaactataag agtagaatct 2220agtaaaccac ctgtatttaa
gaatgatggt aatagtggtg atggtggtaa tactgaatat 2280aattttaatt ttgatttatc
aggattgcaa gatactgggc tttattctgg taaacttaaa 2340tctggtattc gtgtgcaagg
taattacact tacacaggtg ctccatcttt aaatctggtt 2400gtttacagaa ataatagtgt
tgtatccact tttccagtag gttctccttt tgatatcact 2460ataacaacag aaactgataa
ggttatcctt tcattacaac ctcaacatgg gttggcaaca 2520gttactggta ctggcacaat
aacaattcct aatgataaat tagcaattgt ttatgataag 2580ttatttaaat taccacatga
tttagaaaat ataagaatac aagtaaatgc attattcata 2640tcgagtacac aaaatgaatt
agctaaagaa gtaaatgacc atgatattga agaagttgca 2700ttgaaagtag atgcattatc
ggatgaagta tttggaaaag agaaaaaaga attacgtaaa 2760ctggtcaatc aagcgaaacg
tttaagtaaa gcacgaaacc ttctggtagg aggcaatttt 2820gataattggg aagcttggta
taaaggaaaa gaagttgcaa gagtatctga tcatgaatta 2880ttgaagagtg atcatgtatt
attaccgcct ccaactatgt atccatccta tatatatcaa 2940aaagtagaag aaacaaaatt
aaagccaaat actcgttata tgatttctgg tttcatcgca 3000catgcggaag atttagaaat
tgtggtttct cgttatgggc aagaagtaag gaaaatagtg 3060caagttccat atggagaagc
tttcccatta acatccaatg gatcaatttg ttgtacacca 3120agttttagac gtgatggaaa
actatcagat ccacatttct ttagttatag tattgatgta 3180ggtgaactgg atatgacggc
aggtccaggt attgaattgg gacttcgtat tgtagatcga 3240ttaggaatgg cccgtgtaag
taatttagaa attcgtgaag atcgttcttt aacagcaaat 3300gaaatacgaa aagtgcaacg
tatggcaaga aattggagaa ccgaatatga gaaagaacgt 3360gcagaagtaa cagcattaat
tgaacctgta ttaaaccaaa tcaatgcgtt atatgaaaat 3420ggagattgga atggttctat
tcgttcagat atttcgtact acgatataga atctattgta 3480ttaccaacat taccaagatt
acgtcattgg tttgttcctg atatgttaac tgaacatgga 3540aatatcatga atcgattcga
agaagcatta aatcgtgctt atacacagct ggaaggaaat 3600acactattgc ataacggtca
ttttacaaca gatgcggtaa attggatgat acaaggagat 3660gcacatcagg taatattaga
agatggtaga cgtgtattac gattaccaga ctggtcttcg 3720agtgtatccc aaacaattga
aatcgagaaa tttgatccag ataaagaata caacttagta 3780tttcatgcgc aaggagaagg
aacggttacg ttggagcatg gagaaaaaac aaaatatata 3840gaaacgcata cacatcattt
tgcgaatttt acaacatcac aaagtcaagg aattacgttt 3900gaatcgaata aggtgaccgt
ggaaatttct tcagaagatg gggaattatt ggtagatcat 3960atcgcacttg tggaagttcc
tatgtttaac aagaatcaaa tggtcaatga aaatagagat 4020gtaaatataa atagcaatac
aaatatgaat aatagcaata atcaa 4065654772DNABacillus
thuringiensis 65tacccttcat atagtataaa gggacaacct cctatttata tatatattat
tgtaattaca 60ttatagaaga tcaaacagaa aaaagttttt tgaattggtt caatatacca
actatatagc 120ataaactatc tttgtattta tttataataa ttttttaagg agtgaaataa
tgacaacaat 180aaatgaatta tatccggctg taccttataa tgtactggca tatgctccac
cacttaattt 240agctgattcg acaccatggg gtcaaatagt tgttgctgat gcaattaaag
aagcttggga 300taattttcaa aaatatggtg tattagattt aacagctata aatcaagggt
ttgatgatgc 360aaatacaggt tcttttagtt atcaagcttt aatacaaact gttttgggta
ttataggtac 420aatttggtat gacagttcct gtggctgctc catttgcagc tacagcgcct
attattagtt 480tatttgtagg atttttttgg cctaaaaaag ataagggacc acaattaatc
gatataattg 540ataaagaaat taaaaaatta ttagataagg aattaggaga gcaaaaacgt
aatgatttag 600ttagtgcttt aaatgagatg caagagggag caaatgagtt aagtgatatt
atgactaatg 660cactttttga aggtactata cagggaaatg ttgttactaa tgataaccct
caaggtaaaa 720ggcgaactcc taaagctcca acagttagtg attatgagaa tgtttattcg
gcatattttg 780tggaacatgt ggattttaga aacaaaatat ctacgtttct tactggttct
tatgatctta 840tagcactccc attatatgca ttagcaaaaa caatggagct ttcattgtat
caatcattta 900ttaattttgc taataaatgg atggattttg tatatacaaa agcaattaat
gaatcagcaa 960ctgatgatat gaaaagagat tatcaagcga gatacaatac tcaaaaaagt
aatttagctg 1020tacaaaaaac acaattgatt aacaaaatta aagatggtac agatgctgtt
atgaaagttt 1080ttaaagatac caataattta ccttcaatag gtactaataa attagcagta
aatgctcgta 1140ataagtatat tagggcctta caaataaatt gtttagattt agttgctttg
tggcctggct 1200tatatccaga tgaatatctt ttaccattac aattagataa aacacgtgtt
gtattttctg 1260atacaatggg acctgatgaa acacatgatg gtcaaatgaa agttttaaat
atattagact 1320caactacaag ttataaccat caagatatag gaataagtac aactcaagat
gtaaattctt 1380tattatttta tccaagaaaa gaactgttag aattagattt tgctaaatat
atttcatcta 1440gtagtcgttt ttgggtttat ggatttggct taaaatattc agatgataac
ttttatagat 1500atggtgataa cgatccaagc agtgatttta aacctgcata taagtggttt
acgaaaaatt 1560cccagttcga aaaccttcct acttatggaa atcctactcc tattactaat
ttaaatgcta 1620aaactcaagt aacttcttat cttgatgcat taatatatta tatagacgga
ggaactaatc 1680tatataataa tgcgattctt catgatacag ggggttatat tccgggatat
ccaggtgtag 1740aaggatatgg tatgagtaat aatgaacctt tagcaggaca aaaattaaat
gctttatatc 1800ctataaaagt ggaaaatgta agtggttcac aaggaaaatt aggaacaata
gcagcttatg 1860ttcctttaaa tttacaacca gaaaatatta ttggtgatgc tgatccgaat
acaggttttc 1920cccttaatgt aattaaagga tttccatttg aaaaatatgg acctgattat
gagggacgag 1980gaatttcggt tgtaaaagaa tggataaatg gtgcaaatgc tgtaaaattg
tctccaggtc 2040aatcagttgg ggtacaaatt aaaaatataa caaaacaaaa ttatcaaatt
cgtactcgtt 2100atgcaagtaa taacagtaat caagtatatt ttaatgtaga tccaggtgga
tcaccattat 2160ttgcacaatc agtaacattt gaatctacaa caaatgttac aagtggccaa
caaggcgaaa 2220atggtagata tacattaaaa actatttttt ctggtaatga tctacttaca
gtagaaatcc 2280ctgttggaaa tttttatgtg catgttacga ataaaggatc ttctgatatc
tttttagatc 2340gtcttgagtt ttctacagtt ccttcatatg ttatatattc aggtgattat
gatgctacag 2400gtacagatga tgtcttattg tcagatccac atgagtattt ttatgatgtc
atagtgaatg 2460gtactgctag tcattctagt gcagctactt ctatgaattt gctcaataaa
ggaaccgtag 2520taagaagcat tgatattcca ggtcactcaa cgtcttattc tgtacagtat
tcagttccag 2580aaggatttga tgaagttaga attctcagtt ctcttccgga tattagtgga
actataagag 2640tagaatctag taaaccacct gtatttaaga atgatggtaa tagtggtgat
ggtggtaata 2700ctgaatataa ttttaatttt gatttatcag gattgcaaga tactgggctt
tattctggta 2760aacttaaatc tggtattcgt gtgcaaggta attacactta cacaggtgct
ccatctttaa 2820atctggttgt ttacagaaat aatagtgttg tatccacttt tccagtaggt
tctccttttg 2880atatcactat aacaacagaa actgataagg ttatcctttc attacaacct
caacatgggt 2940tggcaacagt tactggtact ggcacaataa caattcctaa tgataaatta
gcaattgttt 3000atgataagtt atttaaatta ccacatgatt tagaaaatat aagaatacaa
gtaaatgcat 3060tattcatatc gagtacacaa aatgaattag ctaaagaagt aaatgaccat
gatattgaag 3120aagttgcatt gaaagtagat gcattatcgg atgaagtatt tggaaaagag
aaaaaagaat 3180tacgtaaact ggtcaatcaa gcgaaacgtt taagtaaagc acgaaacctt
ctggtaggag 3240gcaattttga taattgggaa gcttggtata aaggaaaaga agttgcaaga
gtatctgatc 3300atgaattatt gaagagtgat catgtattat taccgcctcc aactatgtat
ccatcctata 3360tatatcaaaa agtagaagaa acaaaattaa agccaaatac tcgttatatg
atttctggtt 3420tcatcgcaca tgcggaagat ttagaaattg tggtttctcg ttatgggcaa
gaagtaagga 3480aaatagtgca agttccatat ggagaagctt tcccattaac atccaatgga
tcaatttgtt 3540gtacaccaag ttttagacgt gatggaaaac tatcagatcc acatttcttt
agttatagta 3600ttgatgtagg tgaactggat atgacggcag gtccaggtat tgaattggga
cttcgtattg 3660tagatcgatt aggaatggcc cgtgtaagta atttagaaat tcgtgaagat
cgttctttaa 3720cagcaaatga aatacgaaaa gtgcaacgta tggcaagaaa ttggagaacc
gaatatgaga 3780aagaacgtgc agaagtaaca gcattaattg aacctgtatt aaaccaaatc
aatgcgttat 3840atgaaaatgg agattggaat ggttctattc gttcagatat ttcgtactac
gatatagaat 3900ctattgtatt accaacatta ccaagattac gtcattggtt tgttcctgat
atgttaactg 3960aacatggaaa tatcatgaat cgattcgaag aagcattaaa tcgtgcttat
acacagctgg 4020aaggaaatac actattgcat aacggtcatt ttacaacaga tgcggtaaat
tggatgatac 4080aaggagatgc acatcaggta atattagaag atggtagacg tgtattacga
ttaccagact 4140ggtcttcgag tgtatcccaa acaattgaaa tcgagaaatt tgatccagat
aaagaataca 4200acttagtatt tcatgcgcaa ggagaaggaa cggttacgtt ggagcatgga
gaaaaaacaa 4260aatatataga aacgcataca catcattttg cgaattttac aacatcacaa
agtcaaggaa 4320ttacgtttga atcgaataag gtgaccgtgg aaatttcttc agaagatggg
gaattattgg 4380tagatcatat cgcacttgtg gaagttccta tgtttaacaa gaatcaaatg
gtcaatgaaa 4440atagagatgt aaatataaat agcaatacaa atatgaataa tagcaataat
caataacttt 4500tatatgtaaa caggtgcaag tgtttgttgc acctgttttt tcaccctgtt
tctaaaaaaa 4560tgaggaggtt agatgtatgc aaaagaaatc aaaagtaata gaaccaacaa
ataatagtat 4620gtcagtaaat aataatattt cattaaaaac accgatgcca attgggtata
gaaaaaaatc 4680aggttgtggt tgtgggaaac gtcgttaagt ggaaaatgaa tctgtggaat
ggataaaaac 4740aagtgtgaaa gttgaacgtt catccaattt ac
477266723DNABacillus thuringiensis 66atggccgata tgcctgtaaa
taatactcat atatcacaaa atgattttcc aatctgtaca 60actgaagaag aacgtttaat
tccattttgt tgtttggttc agctaccaca tgattttcaa 120cttgttcctt attgcaaacc
acgtcttgta tacaacattg gatgccttgg aacgactaaa 180gaaacgtgta aaaaaaccat
acaagtagag gattgtggac aaacggaaat tgatttgcag 240atattaaagg caaaaggatg
catcaccttt cttgtgaata tagatgtaga acctgtctgt 300gaagaagaga tttgctcgag
tgttccgcac acaaaggaca tgattttgtg ttgtaaagga 360accgtatgtg tagacaaaat
tttaaaatgc agtgttgact gtttacctga tattcattta 420gattgtgaga atgtaaaagt
ttgtgattta caagtaaaat cactctgcga ggaagattgt 480cattctgtaa agattacagg
gtattttcag atttgtattg attaaataaa gatttttaaa 540tgcatacttt tttacatcca
ctcaaataga gagagtgttg caaggttaaa aagcaatatg 600tttttataaa tatttattta
tttcttttgt cctggtgcag aaaaataaaa taatttcaaa 660atgatttttc taactttgcc
ataatactac taaaaaggtg atagagagac taagaatgat 720aag
72367174PRTBacillus
thuringiensis 67Met Ala Asp Met Pro Val Asn Asn Thr His Ile Ser Gln Asn
Asp Phe1 5 10 15
Pro Ile Cys Thr Thr Glu Glu Glu Arg Leu Ile Pro Phe Cys Cys Leu
20 25 30 Val Gln Leu Pro His
Asp Phe Gln Leu Val Pro Tyr Cys Lys Pro Arg 35 40
45 Leu Val Tyr Asn Ile Gly Cys Leu Gly Thr
Thr Lys Glu Thr Cys Lys 50 55 60
Lys Thr Ile Gln Val Glu Asp Cys Gly Gln Thr Glu Ile Asp Leu
Gln65 70 75 80 Ile
Leu Lys Ala Lys Gly Cys Ile Thr Phe Leu Val Asn Ile Asp Val
85 90 95 Glu Pro Val Cys Glu Glu
Glu Ile Cys Ser Ser Val Pro His Thr Lys 100
105 110 Asp Met Ile Leu Cys Cys Lys Gly Thr Val
Cys Val Asp Lys Ile Leu 115 120
125 Lys Cys Ser Val Asp Cys Leu Pro Asp Ile His Leu Asp Cys
Glu Asn 130 135 140
Val Lys Val Cys Asp Leu Gln Val Lys Ser Leu Cys Glu Glu Asp Cys145
150 155 160 His Ser Val Lys Ile
Thr Gly Tyr Phe Gln Ile Cys Ile Asp 165
170 68589DNABacillus thuringiensis 68atgacatttg
attacgagga agaagaaagg gatttgtccc aactccgacc atctgattct 60tcaagatcaa
caggtgaacc aagaacggat tgtgacctaa ccacaaattg tgaagttcct 120ttttgttgcg
tcgtcacgct cccacatgga tttcagtacg aatcccgaaa acaaacaaag 180cttgtatacg
atattagttg tcttaccttt gcacacgaga tgtgtcaacg atctatcaac 240gtggatcagt
gtggtacagt tgatgtagat ttacaagtct taaaaataaa gggctgtgtg 300tcactgtata
tcaacgttcc gattctaccc attcgtgagg agacgatgtg tacgttacac 360agacagccga
catctctcta tacctgttgt caggataccc tttgtgtaga tcatattgtg 420aaatgtagtg
tgggaagtct cccttactac gtgttggatg gcaatcatat acaagtgtgt 480gacttacagg
tacgtcctgt atcagaagta catccacacg tattacaagt atccggtcgt 540tttgagtttt
tatatacgta agtatctata tgaggagtga gataaagat
58969186PRTBacillus thuringiensis 69Met Thr Phe Asp Tyr Glu Glu Glu Glu
Arg Asp Leu Ser Gln Leu Arg1 5 10
15 Pro Ser Asp Ser Ser Arg Ser Thr Gly Glu Pro Arg Thr Asp
Cys Asp 20 25 30
Leu Thr Thr Asn Cys Glu Val Pro Phe Cys Cys Val Val Thr Leu Pro 35
40 45 His Gly Phe Gln Tyr
Glu Ser Arg Lys Gln Thr Lys Leu Val Tyr Asp 50 55
60 Ile Ser Cys Leu Thr Phe Ala His Glu Met
Cys Gln Arg Ser Ile Asn65 70 75
80 Val Asp Gln Cys Gly Thr Val Asp Val Asp Leu Gln Val Leu Lys
Ile 85 90 95 Lys
Gly Cys Val Ser Leu Tyr Ile Asn Val Pro Ile Leu Pro Ile Arg
100 105 110 Glu Glu Thr Met Cys
Thr Leu His Arg Gln Pro Thr Ser Leu Tyr Thr 115
120 125 Cys Cys Gln Asp Thr Leu Cys Val Asp
His Ile Val Lys Cys Ser Val 130 135
140 Gly Ser Leu Pro Tyr Tyr Val Leu Asp Gly Asn His Ile
Gln Val Cys145 150 155
160 Asp Leu Gln Val Arg Pro Val Ser Glu Val His Pro His Val Leu Gln
165 170 175 Val Ser Gly Arg
Phe Glu Phe Leu Tyr Thr 180 185
704PRTArtificial SequenceER targeting peptide 70Lys Asp Glu Leu1
User Contributions:
Comment about this patent or add new information about this topic: