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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。
The HCV NS5 gene fragment was amplified by RT PCR from the sera of patients both positive for both HCV and HCV RNA. The recombinant plasmid pET 2 8a containing this gene fragment was transformed into E. coli BL21 (DE3) to express NS5 protein efficiently. The protein can be purified by immobilized metal ion chelation chromatography (IMAC). The result of Western blot and ELISA showed that the purified NS5 protein could react specifically with the serum of hepatitis C patients, which may have a good application prospect in the diagnosis of HCV infection.