Large-scale production and characterization of Bacillus thuringiensis subsp. tenebrionis insecticidal protein from Escherichia coli.
Appl Microbiol Biotechnol
; 47(3): 255-61, 1997 Mar.
Article
en En
| MEDLINE
| ID: mdl-9114517
Bacillus thuringiensis subsp. tenebrionis insecticidal protein was produced in recombinant Escherichia coli and purified to near homogeneity to provide quantities of protein for safety-assessment studies associated with the registration of transgenic potato plants. The 68-kDa protein is produced naturally by Bacillus thuringiensis subsp. tenebrionis by translation initiation at an internal initiation site in the native DNA sequence. The gene sequence specific for this truncated protein was expressed in E. coli strain JM 101 and fermented at the 1000-1 scale. The protein accumulated as insoluble inclusion bodies, and was purified by extraction at pH 10.8 with carbonate buffer, selective precipitation at pH 9.0, and differential centrifugation. No chromatography steps were required to produce over 50 g purified protein as a lyophilized powder with a purity greater than 95% and demonstrating full insecticidal activity against Colorado potato beetle larvae. The protein was further characterized to assure identity and suitability for use in safety-assessment studies.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Bacillus thuringiensis
/
Proteínas Bacterianas
/
Toxinas Bacterianas
/
Proteínas Recombinantes
/
Control Biológico de Vectores
/
Endotoxinas
/
Escherichia coli
Límite:
Animals
Idioma:
En
Revista:
Appl Microbiol Biotechnol
Año:
1997
Tipo del documento:
Article
País de afiliación:
Estados Unidos
Pais de publicación:
Alemania