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Site-directed mutation of PoIFN-alpha and its expression in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology ; (12): 339-342, 2002.
Artículo en Chino | WPRIM | ID: wpr-231321
ABSTRACT
By using huge primer PCR Cys86 (TGC) of PoIFN-alpha was mutated to Tyr(TAC), and the first code TGT was simultaneously changed to TGC, which is a bias code of E. coli. The expression plasmid pGEX-IFN was constructed successfully. Recombinant porcine IFN alpha, which is expressed as inclusion bodies, was about 20% of the total proteins. The inclusion body was dissolved in 8 mol/L urea and subsequently renatured by dilution in refolding buffer. In order to obtain pure protein, the renatured IFN alpha was purified by FPLC, and the cytokine activity (5200 IU/mg) was verified by inhibiting the cytopathic effect.
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Precursores de Proteínas / Proteínas Recombinantes / Mutagénesis Sitio-Dirigida / Interferón-alfa / Escherichia coli / Genética Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2002 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Precursores de Proteínas / Proteínas Recombinantes / Mutagénesis Sitio-Dirigida / Interferón-alfa / Escherichia coli / Genética Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2002 Tipo del documento: Artículo