丙型肝炎病毒NS_5区基因片段的克隆与表达

CLONING AND EXPRESSION OF NS5 GENE OF HEPATITIS C VIRUS

应用ＲＴ一ＰＣＲ和基因重组技术，从湖南地区ＨＣＶ感染者血清中获得ＨＣＶＮＳ＿５区的基因克隆。经核苷酸序列的比较分析，此克隆片段属１ｂ型ＨＣＶ基因。应用ＰＭＡＬ－ＣＲＩ质粒在大肠杆菌中高效表达了此基因片段，并获融合蛋白ＭＢＰ－ＨＣＶ·ＮＳ＿５，经ＷｅｓｔｅｒｎＢｌｏｔ分析证实该融合蛋白有较特异的ＨＣＶ抗原性，可用于ＨＣＶ感染的临床诊断和发病机制的研究。

gene fragment of NS5 of HCV-Hun was cloned by RT-PCR and gene recombinant tech-niques from blood of HCV infected patients in Hunan province.Comparing cDNA sequences ofour clones with those of HCV-1, HCV-J,HCV-J60,and HCV-J8,it was found that this gene frag-ment of HCV-Hun belonged to genotype 1b of HCV. The fusion protein containing the peptideencoded by HCV· NS5 (MBP-HCV· NS5)was synthesized by E.coli with recombinant plasmids. The specific HCV antigenicity of the fusion protein was identified by Western blotting. This fu-sion protein should be useful in the study on pathogenesis and diagnosis of HCV infection.