HCV NS4基因在大肠杆菌中表达及蛋白质提纯

Expression of the Full-Length HCV NS4 Gene in Escherichia coli and Purification of the Protein

应用 PCR方法扩增出人丙型肝炎病毒全长 N S4基因的 DNA片段 ,克隆入表达载体 p QE3 2并转化E.coli JM10 9,经 IPTG诱导表达出 4 2× 10 3蛋白 .利用 Ni- NTA- agarose纯化得到纯化产物 ,Western- blot鉴定该蛋白能够与抗 NS4的单克隆抗体发生特异性反应 .

A full-length HCV NS4 gene was obtained by PCR method. The DNA fragment was cloned into expressing vector pQE32 and the recombinant plasmid was transformed to E.coli JM109 strain. Induced by IPTG, the NS4 gene was expressed into a protein with molecular weight of 42×10 3 in E.coli. SDS-PAGE proved that the protein was purified to homogenesis by Ni-NTA-agarose column chromatography. Westernblot showed that the protein could react strongly with specific anti-NS4 monoclonal antibody.