Universal and specific probes for Bacillus thuringiensis endotoxin genes
AssigneeMycogen Corporation
InventorsFrank H. Gaertner, August J. Sick, George E. Schwab
Life of this patent
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Abstract
This invention concerns novel gene probes which can be used to identify DNA from Bacillus thuringiensis microbes which encode insecticidally-active protein endotoxins. The invention probes greatly facilitate the search for useful microbes hosting genes which encode insecticidally-active toxins.
Claims36
A gene probe for identifying Bacillus thuringiensis DNA encoding insecticidally-active protein endotoxins consisting essentially of a nucleotide sequence selected from the group consisting of
- (a) GGACCAGGATTTACAGGAGGAGAT;
- (b) ATGATTCATGCGGCAGATA;
- (c) GGTGATTTTACACAAGGGGTAATGGGGTGGCATG;
- (d) AAAGCTCTTGCAGAGTTACAGGG;
- (e) GAATTATACTTGGTTCAGGCCC;
- (f) GTCCAATCGGAAAATGTGCCC;
- (g) CCAATTCACTTCCCATCGAC;
- (h) GCTGAAGAACTTCCTATTCGTGGTGGTGAGC;
- (i) CGTTATGGAGAGCAGCGCA;
- (j) GTTAGCGCATCAACAGTCCAAACGGG;
- (k) CATGTTTATACTAACCATTGTGTGGATACG;
- (l) CGTAGCAGCAGAAATCGGCTTGGGC;
- (m) GGGAGGAACAAATATGAATCCTTATC;
- (n) CAGGCTATCCGTTAGCGAATGACTTACAAGGG;
- (o) AATGTTGGCGCGGTCAGCTGGG;
- and
- (p) TACAATGGCTATTTAGGTGCACAG.
A process for detecting the presence of genes, or fragments thereof, encoding a protein endotoxin active against insects selected from the group consisting of coleopteran, dipteran, and lepidopteran insects which comprises:
- (a) contacting Bacillus thuringiensis DNA with restriction endonucleases to obtain DNA fragments;
- (b) probing said DNA fragments under hybridizing conditions with a detectably labeled gene probe comprising DNA having the following sequence:
- GGACCAGGATTTACAGGAGGAGAT;
- and
- (c) detecting probe DNA which has hybridized to said Bacillus thuringiensis DNA fragments.
A process for detecting the presence of genes, or fragments thereof, encoding a protein endotoxin active against insects of the order lepidoptera, which comprises:
- (a) contacting Bacillus thuringiensis DNA with restriction endonucleases to obtain DNA fragments;
- (b) probing said DNA fragments under hybridizing conditions with a detectably labeled gene probe comprising DNA having a nucleotide sequence selected from the group consisting of:
- ATGATTCATGCGGCAGATA,
- GAATTATACTTGGTTCAGGCCC,
- GTCCAATCGGAAAATGTGCCC,
- CCAATTCACTTCCCATCGAC,
- GCTGAAGAACTTCCTATTCGTGGTGGTGAGC,
- CGTTATGGAGAGCAGCGCA,
- GTTAGCGCATCAACAGTCCAAACGGG,
- CATGTTTATACTAACCATTGTGTGGATACG, and
- CGTAGCAGCAGAAATCGGCTTGGGC;
- and
- (c) detecting probe DNA which has hybridized to said Bacillus thuringiensis DNA fragments.
A process for detecting the presence of genes, or fragments thereof, encoding a protein endotoxin active against insects of the order diptera, which comprises:
- (a) contacting Bacillus thuringiensis DNA with restriction endonucleases to obtain DNA fragments;
- (b) probing said DNA fragments under hybridizing conditions with a gene probe comprising DNA having a nucleotide sequence selected from the group consisting of:
- GGTGATTTTACACAAGGGGTAATGGGGTGGCATG,
- GGGAGGAACAAATATGAATCCTTATC, and
- CAGGCTATCCGTTAGCGAATGACTTACAAGGG;
- and
- (c) detecting probe DNA which has hybridized to said Bacillus thuringiensis DNA fragments.
A process for detecting the presence of genes, or fragments thereof, encoding a protein endotoxin active against insects of the order coleoptera, which comprises:
- (a) contacting Bacillus thuringiensis DNA with restriction endonucleases to obtain DNA fragments;
- (b) probing said DNA fragments under hybridizing conditions with a gene probe comprising DNA having a nucleotide sequence selected from the group consisting of:
- AAAGCTCTTGCAGAGTTACAGGG,
- AATGTTGGCGCGGTCAGCTGGG, and
- TACAATGGCTATTTAGGTGCACAG;
- and
- (c) detecting probe DNA which has hybridized to said Bacillus thuringiensis DNA fragments.
Dependencies are inferred from the claim text.