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Cloning and characterization of a novel gene cry9Ec1 encoding lepidopteran-specific parasporal inclusion protein from a Bacillus thuringiensis serovar galleriae strain

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A novel -endotoxin gene from a lepidopteran-specific Bacillus thuringiensis serovar galleriae strain was cloned, and the full sequence of the cry gene was determined. The cloned 6.5-kb DNA fragment included the full sequence of the cry gene and three open reading frames located upstream of the cry gene. The gene, designated cry9Ec1, encodes a polypeptide of 1154 amino acid residues with a predicted molecular weight of 130 237. The deduced amino acid sequence of the Cry9Ec1 protein had the highest homology (77.7%) with the Cry9Ea1 protein when compared with existing Cry proteins. The expression, in an acrystalliferous B. thuringiensis strain, of the cry9Ec1 gene was high when controlled by the cyt1A2 promoter, leading to the formation of large spherical inclusions. The purified crystals from the recombinant strain were toxic when tested against two lepidopteran species, Bombyx mori and Plutella xylostella. However, the Cry9Ec1 protein gave no toxicity against Spodoptera litura, Spodoptera exigua, Plodia interpunctella, Helicoverpa zea, and Culex pipiens molestus.Key words: Bacillus thuringiensis serovar galleriae, Cry9Ec1 protein, lepidopteran-specific -endotoxin, Plutella xylostella.

Nous avons cloné un nouveau gène de -endotoxine issu d'une souche de Bacillus thuringiensis sérovar galleriae spécifique aux lépidoptères et avons séquencé le gène cry au complet. Le fragment cloné de 6,5 kb d'ADN renfermait la séquence complète du gène cry et trois cadres de lecture ouverts situés en aval du gène cry. Le gène, dénommé cry9Ec1, code un polypeptide de 1154 acides aminés ayant un poids moléculaire prédit de 130 137. La séquence d'acides aminés déduite de la protéine Cry9Ec1 démontrait une homologie la plus élevée (77,7 %) avec la protéine Cry9Ea1 lorsque comparée à des protéines Cry existantes. L'expression du gène Cry9Ec1 dans une souche de B. thuringiensis acrysallifère était élevée lorsque contrôlée par le promoteur cyt1A2, menant à la formation d'inclusions sphériques volumineuses. Les cristaux purifiés de la souche recombinante étaient toxiques lorsque testés sur deux espèces de lépidoptères, Bombyx mori et Plutella xylostella. Toutefois, la protéine Cry9Ec1 n'a pas démontré de toxicité contre Spodoptera litura, Spodoptera exigua, Plodia interpunctella, Helicoverpa zea et Culex pipiens molestus.Mots clés : Bacillus thuringiensis sérovar galleriae, protéine Cry9Ec1, -endotoxine spécifique aux lépidoptères, Plutella xylostella.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: November 1, 2005

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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