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Chinese Journal of Biotechnology ; (12): 483-486, 2007.
Artigo em Chinês | WPRIM | ID: wpr-328000

RESUMO

Methylotrophic yeast, Pichia pastoris was used to express recombinant batroxobin, and a technology route of producing recombinant protein was finally established. We synthesized batroxobin gene artificially by means of recursive PCR. pPIC9-batroxobin was constructed and transformed into Pichia pastoris GS115 (his4). Recombinant batroxobin was expressed in yeast engineering strain and it was purified from the culture supernatant. 10 mg of recombinant batroxobin was purified from 1 liter fermentation media, it exhibited specific activity of 238 NIH units/mg and had molecular weight of 30.55 kD. The purified recombinant protein converted fibrinogen into fibrin clot in vitro, and shortened bleeding time in vivo. This study laid a foundation of development of hemostatic of recombinant snake venom thrombin-like enzyme.


Assuntos
Animais , Masculino , Camundongos , Batroxobina , Genética , Metabolismo , Farmacologia , Eletroforese em Gel de Poliacrilamida , Expressão Gênica , Hemorragia , Concentração de Íons de Hidrogênio , Pichia , Genética , Reação em Cadeia da Polimerase , Proteínas Recombinantes , Metabolismo , Farmacologia , Fatores de Tempo
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