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.
Article
in Chinese
| 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Protein Precursors
/
Recombinant Proteins
/
Mutagenesis, Site-Directed
/
Interferon-alpha
/
Escherichia coli
/
Genetics
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2002
Type:
Article
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