Polynucleotide compositions encoding broad-spectrum delta endotoxins
AssigneeMonsanto Technology LLC
InventorsThomas Malvar, Amy Jelen Gilmer
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Abstract
Disclosed are novel synthetically-modified B. thuringiensis chimeric crystal proteins having improved insecticidal activity against coleopteran, dipteran and lepidopteran insects. Also disclosed are the nucleic acid segments encoding these novel peptides. Methods of making and using these genes and proteins are disclosed as well as methods for the recombinant expression, and transformation of suitable host cells. Transformed host cells and transgenic plants expressing the modified endotoxin are also aspects of the invention.
Claims78
An isolated and purified Bacillus thuringiensis crystal protein comprising the amino acid sequence of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:26 SEQ ID NO:28, or SEQ ID NO:30.
A method of using a DNA segment that encodes a crystal protein or peptide, comprising the steps of:
- (a) preparing a recombinant vector in which a crystal protein or peptide-encoding DNA segment is positioned under the control of a promoter;
- (b) introducing said recombinant vector into a host cell;
- (c) culturing said host cell under conditions effective to allow expression of the encoded crystal protein or peptide;
- and
- (d) collecting said expressed crystal protein or peptide.
A peptide composition, comprising a B. thuringiensis crystal protein having the amino acid sequence of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:26, SEQ ID NO:28, or SEQ ID NO:30.
A purified antibody that binds to a B. thuringiensis crystal protein having the amino acid sequence of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:26, SEQ ID NO:28, or SEQ ID NO:30.
A transgenic plant having incorporated into its genome a transgene that encodes a B. thuringiensis crystal protein having the amino sequence of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:26, SEQ ID NO:28, or SEQ ID NO:30.
A Bacillus thuringiensis cell that produces a crystal protein comprising the amino acid sequence of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:26, SEQ ID NO:28, or SEQ ID NO:30.
A Bacillus thuringiensis cell that produces a crystal protein encoded by the nucleic acid sequence of SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:25, SEQ ID NO:27, or SEQ ID NO:29.
A Bacillus thuringiensis cell having the NRRL accession number B-21579.
A Bacillus thuringiensis cell having the NRRL accession number B-21580.
A Bacillus thuringiensis cell having the NRRL accession number B-21581.
A Bacillus thuringiensis cell having the NRRL accession number B-21635.
A Bacillus thuringiensis cell having the NRRL accession number B-21636.
A Bacillus thuringiensis cell having the designation EG11090.
A composition comprising from about 0.5% to about 99% by weight of a crystal protein having the amino acid sequence of SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:26, SEQ ID NO:28, or SEQ ID NO:30.
A composition comprising from about 0.5% to about 99% by weight of a crystal protein encoded by the nucleic acid sequence of SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:25, SEQ ID NO:27, or SEQ ID NO:29.
A composition comprising a B. thuringiensis crystal protein prepared by a process comprising the steps of:
- (a) culturing a Bacillus thuringiensis EG11090, NRRL B-21579, NRRL B-21580, NRRL B-21581, NRRL B-21635, or NRRL B-21636 cell under conditions effective to produce a B. thuringiensis crystal protein;
- and
- (b) obtaining said B. thuringiensis crystal protein from said cell.
A method of preparing a B. thuringiensis crystal protein comprising:
- (a) culturing a Bacillus thuringiensis EG11090, NRRL B-21579, NRRL B-21580, NRRL B-21581, NRRL B-21635, or NRRL B-21636 cell under conditions effective to produce a B. thuringiensis crystal protein;
- and
- (b) obtaining said B. thuringiensis crystal protein from said cell.
Dependencies are inferred from the claim text.