Chemistry: molecular biology and microbiology – Plant cell or cell line – per se ; composition thereof;... – Plant cell or cell line – per se – is pest or herbicide...
Reexamination Certificate
2001-04-19
2004-05-18
Bugaisky, Gabriele (Department: 1653)
Chemistry: molecular biology and microbiology
Plant cell or cell line, per se ; composition thereof;...
Plant cell or cell line, per se, is pest or herbicide...
C435S419000, C435S252300, C435S254110, C435S320100, C424S093200, C424S093461, C536S023710
Reexamination Certificate
active
06737273
ABSTRACT:
BACKGROUND OF THE INVENTION
The most widely used microbial pesticides are derived from the bacterium
Bacillus thuringiensis
. This bacterial agent is used to control a wide range of leaf-eating caterpillars and beetles, as well as mosquitos.
Bacillus thuringiensis
produces a proteinaceous parasporal body or crystal which is toxic upon ingestion by a susceptible insect host. For example,
B. thuringiensis
subsp.
kurstaki
HD-1 produces a crystal inclusion consisting of a biotoxin called a delta toxin which is toxic to the larvae of a number of lepidopteran insects. The cloning, sequencing, and expression of this
B.t
. crystal protein gene in
Escherichia coli
has been described in the published literature (Schnepf, H. E. and Whitely, H. R. [1981] Proc. Natl. Acad. Sci. USA 78:2893-2897; Schnepf et al.). U.S. Pat. No. 4,448,885 and U.S. Pat. No. 4,467,036 both disclose the expression of
B.t
. crystal protein in
E. coli.
BRIEF SUMMARY OF THE INVENTION
The subject invention concerns a novel
Bacillus thuringiensis
isolate designated
B.t
. PS81I which has activity against all lepidopteran pests tested.
Also disclosed and claimed are novel toxin genes which express toxins toxic to lepidopteran insects. These toxin genes can be transferred to suitable hosts via a plasmid vector.
Specifically, the invention comprises the novel
B.t
. isolate denoted
B.t
. PS81I, mutants thereof, and novel &dgr;-endotoxin genes derived from this
B.t
. isolate which encode proteins which are active against lepidopteran pests.
BRIEF DESCRIPTION OF THE SEQUENCES
SEQ ID NO:1 is the nucleotide sequence of the novel
B.t
. toxin gene PS81I2.
SEQ ID NO:2 is the amino acid sequence of the novel
B.t
. toxin PS81IA2.
SEQ ID NO:3 is the nucleotide sequence of the novel
B.t
. toxin gene PS81B.
SEQ ID NO:4 is the amino acid sequence of the novel
B.t
. toxin PS81B.
SEQ ID NO:5 is the nucleotide sequence of the novel
B.t
. toxin gene PS81IB2.
SEQ ID NO:6 is the amino acid sequence of the novel
B.t
. toxin PS81IB2.
SEQ ID NO:7 is the nucleotide sequence of the novel
B.t
. toxin gene PS81IA.
SEQ ID NO:8 is the amino acid sequence of the novel
B.t
. toxin PS81IA.
SEQ ID NO:9 is the 42-mer oligonucleotide constructed to the sequence of the insert in pM2,31-4.
SEQ ID NO:10 is the 40-mer oligonucleotide constructed to the sequence of the insert in pM2,31-1.
REFERENCES:
patent: 5126133 (1992-06-01), Payne et al.
patent: 5164180 (1992-11-01), Payne et al.
patent: 5188960 (1993-02-01), Payne et al.
patent: 5206166 (1993-04-01), Payne et al.
patent: 5246852 (1993-09-01), Payne et al.
patent: 5407825 (1995-04-01), Payne et al.
patent: 5691308 (1997-11-01), Payne et al.
patent: 6096708 (2000-08-01), Payne et al.
patent: 0632335 (1993-05-01), None
patent: 0 400 246 (1990-05-01), None
Sutton et al. Synthetic crylllA gene fromBacillus thuringiensisimproved for high expression in plants. 1992. Transgenic Research vol. 1(5) pp. 228-236.*
Honee et al., “Nucleotide sequence of crystal protein gene isolated fromB. thuringensissubspeciesentomocidus60.5 coding for a toxin highly active againstSpodopteraspecies,”Nucleic Acids Research(1988), p. 6240, vol. 16, Issue 13.
Sanchis et al., “Nucleotide sequence and analysis of the N-terminal coding region of theSpodoptera-active &dgr;-endotoxin gene ofBacillus thuringiensis aizawai7.29,”Molecular Microbiology, 1989, pp. 229-238, vol. 3, Issue 2.
Hofte et al., “Nucleotide sequence and deduced amino acid sequence of a new Lepidoptera-specific crystal protein gene fromBacillus thuringiensis, ”Nucleic Acids Research, 1990, p. 5545, vol. 18.
Chambers et al., “Isolation and Characterization of a Novel Insecticidal Crystal Protein Gene fromBacillus thuringiensissubsp.aizawai,”Journal of Bacteriology, 1991, pp. 3966-3976, vol. 173, Issue 13.
Payne Jewel
Sick August J.
Bugaisky Gabriele
Myocogen Corporation
Saliwanchik Lloyd & Saliwanchik
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