丙型肝炎病毒非结构区NS5基因片段的克隆和表达

从抗 HCV和HCVRNA均为阳性的病人血清中 ,采用RT PCR技术扩增得到HCVNS5基因片段。用含此基因片段的重组质粒pET 2 8a转化大肠杆菌BL2 1(DE3)后可高效表达NS5蛋白。该蛋白可用固定化金属离子螯合层析法 (IMAC)纯化。经Westernblot和ELISA检测结果表明 ,纯化的NS5蛋白可与丙型肝炎患者血清发生特异反应 ,在HCV感染的诊断中可能具有良好的应用前景。

Cloning and Expression of NS5 Gene Fragment of Hepatitis C Virus

The NS5 gene fragment of HCV was amplified from anti HCV and HCV RNA positive sera by RT PCR.After the gene fragment was inserted into expression vector pET 28a and transformed to E.coli BL21(DE3),NS5 protein was successfully expressed and purified by immobilized metal affinity chromography(IMAC).Western blot and ELISA proved that the NS5 protein was specifically reacted with the sera of patients with hepatitis C and might be applied for the diagnosis of HCV infection.