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AssigneeMonsanto Technology LLC
InventorsJames Baum, Judith Donovan, William Donovan, James Engleman, Karina Krasomil-Osterfeld, John Pitkin, James Roberts
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The present invention relates to the isolation and characterization of nucleotide sequences encoding novel insecticidal proteins secreted into the extracellular space from Bacillus thuringiensis and related strains. The proteins are isolated from culture supernatants of Bacillus thuringiensis and related strains and display insecticidal activity against coleopteran insects including Colorado potato beetle (Lymantria dispar) and Southern Corn Rootworm (Diabrotica undecempunctata). Insecticidal proteins encoded by nucleotide sequences that hybridize to the isolated and characterized nucleotide sequences are disclosed. Also disclosed are methods of making and using transgenic cells and plants comprising the novel nucleotide sequence of the invention.
An isolated polynucleotide which encodes a Bacillus thuringiensis insecticidal toxin or insecticidal fragment thereof, wherein said polynucleotide hybridizes under conditions selected from the group consisting of stringent hybridization conditions and specific hybridization conditions with one or more of the nucleotide sequences selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3 (tic901), SEQ ID NO:5 (tic1201), SEQ ID NO:7 (tic407), SEQ ID NO:9 (tic417), SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:32 or with the complement thereof.
A polynucleotide comprising a nucleotide sequence which encodes an approximately 34 to about 39 kDa toxin active against a coleopteran pest, wherein said nucleotide sequence has been optimized for expression in plants, and wherein said toxin comprises the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:31, and SEQ ID NO:33.
A host cell transformed to contain a polynucleotide encoding an insecticidal protein or insecticidal fragment thereof wherein said polynucleotide comprises a nucleotide sequence as set forth in a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, SEQ ED NO:7, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:30, and SEQ ID NO:32.
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A method for detecting a first nucleotide sequence encoding an insecticidal protein wherein said first nucleotide sequence hybridizes to a second nucleotide sequence that is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3 (tic901), SEQ ID NO:5 (tic1201), SEQ ID NO:7 (tic407), SEQ ID NO:9 (tic417), SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:32 (tic431) or with the complement thereof under stringent hybridization conditions.
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A method for detecting a first nucleotide sequence encoding an insecticidal protein, comprising the steps of (a) producing an amplicon from a template comprising said first nucleotide sequence in a sample using amplification primers comprising the sequences as set forth in SEQ ID NO:13 and SEQ ID NO:14;
An isolated nucleic acid molecule comprising a polynucleotide sequence encoding a toxin protein wherein said toxin protein comprises a sequence that exhibits at least about 70% sequence identity to a nucleotide sequence selected from the group of amino acid sequences consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:31, and SEQ ID NO:33, or a coleopteran-active variant or portion thereof.
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A recombinant DNA construct, comprising a polynucleotide sequence encoding an insecticidal protein, wherein said polynucleotide sequence is at least about 70% identical to SEQ ID NO:13, said insecticidal protein being selected from the group consisting of all or an insecticidal fragment of a protein as set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:6, SEQ ID NO:10, SEQ ID NO:31, and SEQ ID NO:33.
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Dependencies are inferred from the claim text.