Polynucleotides encoding a 130 kDa pesticidal protein from Bacillus thuringiensis isolate PS201T6
AssigneeMycogen Corporation
InventorsJewel Payne, Kenneth E. Narva, Kendrick Akira Uyeda, Christine Julie Stalder, Tracy Ellis Michaels
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Abstract
Disclosed and claimed are approximately 130 kDa toxins produced by Bacillus thuringiensis PS201T6, which can be used to control pests. Further, claimed are novel genes encoding these .delta.-endotoxins, which can be expressed in other hosts. Expression of the .delta.-endotoxins in such hosts results in the control of susceptible insect pests in the environment of such hosts results in the control of susceptible insect pests in the environment of such hosts.
Claims28
An isolated polynucleotide that encodes a .delta.-endotoxin active against a coleopteran pest, wherein said polynucleotide hybridizes with a 285 bp probe obtainable by PCR amplification, with the primer shown in SEQ ID NO:
- 2 and with the reverse primer shown in SEQ ID NO:
- 3, of cellular DNA from E. coli NM522 having Accession No. NRRL B-21018;
- wherein hybridization occurs at 42.degree. C. in 50% formamide, 5X SSPE, 5X Denhardt's solution, 2% SDS, with 100 .mu.g/ml single stranded DNA;
- and wash occurs at 42.degree. C. in 2X SSC and 0.1% SDS.
An isolated polynucleotide that encodes the .delta.-endotoxin produced by E. coli NM522 having Accession No. NRRL B-21018, or a portion of said .delta.-endotoxin wherein said portion is active against a coleopteran pest.
A plant cell that expresses an isolated polynucleotide that encodes a .delta.-endotoxin, active against a coleopteran pest, wherein said polynucleotide hybridizes with a 285 bp probe obtainable by PCR amplification, with the primer shown in SEQ ID NO:
- 2 and with the reverse primer shown in SEQ ID NO:
- 3, of cellular DNA from E. coli NM522 having Accession No. NRRL B-21018;
- wherein hybridization occurs at 42.degree. C. in 50% formamide, 5X SSPE, 5X Denhardt's solution, 2% SDS, with 100 .mu.g/ml single stranded DNA;
- and wash occurs at 42.degree. C. in 2X SSC and 0.1% SDS.
A plant cell that expresses a polynucleotide that encodes the .delta.-endotoxin produced by E. coli NM522 having Accession No. NRRL B-21018, or a portion of said .delta.-endotoxin wherein said portion is active against a coleopteran pest.
A microbial cell that expresses an isolated polynucleotide that encodes a .delta.-endotoxin, active against a coleopteran pest, wherein said polynucleotide hybridizes with a 285 bp probe obtainable by PCR amplification, with the primer shown in SEQ ID NO:
- 2 and with the reverse primer shown in SEQ ID NO:
- 3, of cellular DNA from E. coli NM522 having Accession No. NRRL B-21018;
- wherein hybridization occurs at 42.degree. C. in 50% formamide, 5X SSPE, 5X Denhardt's solution, 2% SDS, with 100 .mu.g/ml single stranded DNA;
- and wash occurs at 42.degree. C. in 2X SSC and 0.1% SDS.
A transfer vector comprising a polynucleotide that encodes a .delta.-endotoxin, active against a coleopteran pest, wherein said polynucleotide hybridizes with a 285 bp probe obtainable by PCR amplification, with the primer shown in SEQ ID NO:
- 2 and with the reverse primer shown in SEQ ID NO:
- 3, of cellular DNA from E. coli NM522 having Accession No. NRRL B-21018;
- wherein hybridization occurs at 42.degree. C. in 50% formamide, 5X SSPE, 5X Denhardt's solution, 2% SDS, with 100 .mu.g/ml single stranded DNA;
- and wash occurs at 42.degree. C. in 2X SSC and 0.1% SDS.
Dependencies are inferred from the claim text.