Secreted expression of porcine interferon-gamma gene in Pichia pastoris / 生物工程学报
Chinese Journal of Biotechnology
; (12): 731-736, 2005.
Article
en Zh
| WPRIM
| ID: wpr-237082
Biblioteca responsable:
WPRO
ABSTRACT
The porcine interferon-gamma (PoIFN-gamma) gene, in which the sequence encoding signal peptide was replaced by that of the alpha-factor of Saccharomyces cerevisiae, was cloned into Pichia pastoris expression vector pPIC9K. The recombinant plasmid pPIC9K-alpha-PoIFN-gamma was then transformed into Pichia pastoris GS115 cells by electroporation and stable multicopy recombinant Pichia pastoris strains were selected by G418 resistance. Two recombinants of multiple inserts were obtained. SDS-PAGE and Western blot assays of culture broth from a methanol-induced expression strain demonstrated that recombinant PoIFN-gamma, 17kD and 23kD proteins, were secreted into the culture medium. Target proteins, 60% of total proteins, were obtained in the culture medium at the concentration of 108 mg/L. This is the first secreted expression of porcine interferon-gamma gene in Pichia pastoris.
Texto completo:
1
Banco de datos:
WPRIM
Asunto principal:
Pichia
/
Porcinos
/
Proteínas Recombinantes
/
Interferón gamma
/
Electroporación
/
Genética
/
Metabolismo
Límite:
Animals
Idioma:
Zh
Año:
2005
Tipo del documento:
Article