Dna encoding lepidopteran-active delta-endotoxins and its use

Details Dna encoding lepidopteran-active delta-endotoxins and its use Dna encoding lepidopteran-active delta-endotoxins and its use

C - Chemistry, Metallurgy

12

N

C12N 15/32 (2006.01) A01H 5/10 (2006.01) A01N 63/00 (2006.01) C07K 14/325 (2006.01) C07K 19/00 (2006.01) C12N 7/01 (2006.01) C12N 15/82 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2272847

Disclosed are novel synthetically-modified B. thuringiensis nucleic acid segments encoding .delta.-endotoxins having insecticidal activity against lepidopteran insects. Also disclosed are synthetic crystal proteins encoded by these novel nucleic acid sequences. Methods of making and using these genes and proteins are disclosed as well as methods for the recombinant expression, and transformation of suitable host cells. Transformed host cells and transgenic plants expressing the modified endotoxin are also aspects of the invention. Also disclosed are methods for modifying, altering, and mutagenizing specific loop regions between the .alpha. helices in domain 1 of these crystal proteins, including Cry1C, to produce genetically-engineered recombinant cry* genes, and the proteins they encode which have improved insecticidal activity. In preferred embodiments, novel Cry1C* amino acid segments and the modified cry1C* nucleic acid sequences which encode them are disclosed.

Cette invention concerne de nouveaux segments d'acide nucléique B. thuringiensis modifiés synthétiquement, lesquels codent des .delta.-endotoxines ayant une action insecticide contre les insectes de la famille des lépidoptères. Cette invention concerne également des protéines de cristaux synthétiques codées par ces nouvelles séquences d'acide nucléique. Cette invention concerne en outre des procédés de production et d'utilisation de ces gènes et de ces protéines, ainsi que des procédés d'expression recombinante et de transformation de cellules hôtes acceptables. Sous certains aspects, cette invention concerne aussi des cellules hôtes transformées ainsi que des plantes transgéniques exprimant l'endotoxine modifiée. Cette invention concerne aussi des procédés de modification, d'altération et de mutagénisation de zones de boucles spécifiques entre les hélices .alpha. dans le domaine 1 de ces protéines de cristaux, y compris le Cry1C. Ces procédés permettent de produire par génie génétique des gènes cry* recombinants ainsi que les protéines qu'ils codent, lesquelles possèdent une meilleure action insecticide. Dans des modes de réalisation préférés, cette invention offre de nouveaux segments d'acide aminé Cry1C*, ainsi que des séquences d'acide nucléique cry1C* modifiées qui codent ces segments.

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