Bacillus thuringiensis strains and their insecticidal proteins

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4351723, 4353201, 435419, 536 2371, 800250, 800DIG56, A01H 500, A01H 510, C12N 514, C12N 1532, C12N 1582

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058615431

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BRIEF SUMMARY
BACKGROUND OF THE INVENTION

1. Field of the Invention
This invention relates to four novel strains of Bacillus thuringiensis (the "BTS02617A strain", the "BTS02618A strain", the "BTS02654B strain" and the "BTS02652E strain"), each of which produces crystallized proteins (the "BTS02617A crystal proteins", the "BTS02618A crystal proteins", the "BTS02654B crystal proteins" and the "BTS02652E crystal proteins", respectively) which are packaged in crystals (the "BTS02617A crystals", the "BTS02618A crystals", the "BTS02654B crystals" and the "BTS02652E crystals", respectively) during sporulation. The BTS02617A, BTS02618A, BTS02654B and BTS02652E strains were deposited under the provisions of the Budapest Treaty at the Belgian Coordinated Collections of Microorganisms-Collection Laboratorium voor Microbiologie Belgium ("BCCM-LMG"), R.U.G., K. Ledeganckstraat 35, B-9000 Gent.
This invention also relates to an insecticide composition that is active against Lepidoptera and that comprises the BTS02617A, BTS02618A, BTS02654B or BTS02652E strain, as such, or preferably the BTS02617A, BTS02618A, BTS02654B or BTS02652E crystals, crystal proteins or the active component(s) thereof as an active ingredient.
This invention further relates to a gene (the "bTS02618A gene"), which is present in the genome of the BTS02617A, BTS02618A, BTS02654B and BTS02652E strains and which encodes an insecticidal protein (the "BTS02618A protoxin") that is found in the BTS02617A, BTS02618A, BTS02654B and BTS02652E crystals. The BTS02618A protoxin is the protein that is produced by the BTS02617A, BTS02618A, BTS02654B and BTS02652E strains before being packaged into their respective BTS02617A, BTS02618A, BTS02654B and BTS02652E crystals.
This invention still further relates to a toxin (the "BTS02618A toxin") which can be obtained (e.g., by trypsin digestion) from the BTS02618A protoxin. The BTS02618A toxin is an insecticidally active protein which can be liberated from the BTS02617A crystals, the BTS02618A crystals, the BTS02654B crystals, and the BTS02652E crystals, which are produced by the BTS02617A strain, the BTS02618A strain, the BTS02654B strain and the BTS02652E strain, respectively. This toxin and its protoxin have a high activity against a wide range of lepidopteran insects, particularly against Noctuidae, especially against Spodoptera and Aqrotis spp., but also against other important lepidopteran insects such as Pyralidae, particularly the European corn borer, Ostrinia nubilalis, Gelechiidae such as Phthorimaea operculella and Yponomeutidae such as Plutella xylostella. Furthermore, the BTS02618A protein is the first Bt protein with significant activity towards Acrotis seqetum. This new characteristic of the BTS02618A protoxin and toxin ("(pro)toxin"), i.e., the combination of activity against different economically important Lepidopteran insect families such as Noctuidae, Yponomeutidae, Gelechiidae and Pyralidae, makes this (pro)toxin an ideally suited compound for combatting a wide range of insect pests by contacting these insects with the (pro)toxin, e.g., by spraying or by expressing the bTS02618A gene in plant-associated bacteria or in plants. The BTS02618A toxin is believed to represent the smallest portion of the BTS02618A protoxin which is insecticidally effective against Lepidoptera.
This invention also relates to transformed Bacillus thuringiensis strains, containing DNA sequences encoding a BTS02618A protein or variants thereof having substantially the same insecticidal activity.
This invention yet further relates to a chimeric gene that can be used to transform a plant cell and that contains the following operably linked DNA fragments: gene part") encoding an insecticidally effective portion of the BTS02618A protoxin, preferably a truncated part of the bTS02618A gene (the "truncated bTS02618A gene") encoding just the BTS02618A toxin; bTS02618A gene part in a plant cell; and expressing the insecticidally effective bTS02618A gene part in a plant cell. chimeric gene".
This invention also relates to: cotton, the genome of whi

REFERENCES:
patent: 5066667 (1991-11-01), Ehrenfreund et al.
Gleave et al., J. General Microbiology, 138, 55-62 (1992).
Smulevitch et al., Febs Letters, 293, 25-37 (1991).
Vaeck et al., Nature, 328, 33-37 (1987).
Hofte et al., Microbiological Reviews, 53, 242-255 (1989).
Ge et al., J. Biological Chemistry, 266, 17954-17958 (1991).
Reynaerts et al., Med. Fac. Landbouww. Rijksuniv. Gent., 51/3b, 1173-1178 (1986).
Perlak FJ, et al. "Modification of the coding sequence enhances plant expression of insect control protein genes." PNAS 88: 3324-3328, Apr. 1991 .

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