人α1/158Ⅴ型(α1c型)干扰素基因在大肠杆菌中的表达及其产物的初步纯化

EXPRSSION OF INTERFERON-α1/158V(α1c)IN E.coli AND PRELIMINARY PURIFICATION OF EXPRESSION PRODUCT

采用DNA聚合酶链反应(PCR)技术,将本实验室从中国人胎肝细胞染色体DNA中发现和分离的IFN-α1/158V基因的原始克隆,改造成适于进行非融合蛋白原核表达的结构形式,并在大肠杆菌中获得高效表达。测得重组IFN-α1/158V的抗病毒活性为1.9×10~7单位/升菌液。随后又采用以单克隆抗体亲和层析为主的纯化流程对表达产物进行初步纯化,获得了在SDS-聚丙烯酰胺凝胶电泳上呈现单一条带的纯化产物。

IFN-α1/158V ( i.e.IFN-α1c ) , which was recently identified and isolated in our laboratory, was engineered via polymerase chain reaction ( PCR ) to obtain a form that could be expressed as a nonfusion protein in prokaryotic cells. Expression with high efficiency was obtained in E.coli and the recombinant IFN-α1/158V possessed antiviral activity as high as l.9×107U/L culture. A preliminary purification procedure consisting of affinity chromatography was then performed. The purified product appeared as a single band on SDS-PAGE.