丙型肝炎病毒包膜蛋白基因片段的克隆与表达

Cloning and Expression of E1 and E2/NS1 Genes of Hepatitis C Virus

应用ＲＴ－ＰＣＲ和基因重组技术，从湖南地区丙型肝炎病毒（ＨＣＶ）感染者血清中克隆了ＨＣＶ包膜蛋白基因片段（Ｅ１，Ｅ２／ＮＳ１），经与已知基因型的ＨＣＶ－１，ＨＣＶ－Ｊ，ＨＣＶ－Ｊ６和ＨＣＶ－Ｊ８进行核苷酸序列的比较分析，这些基因片段属１ｂ型ＨＣＶ基因。将克隆基因重组于表达质粒ＰＭＡＬ－ＣＲＩ中，在大肠杆菌内进行高效表达，获得了融合蛋白ＭＢＰ－Ｅ１，ＭＢＰ－Ｅ２／ＮＳ１，经ＷｅｓｔｅｒｎＢｌｏｔ分析证实这些融合蛋白具有ＨＣＶ的特异抗原活性，提示这些融合蛋白可用于ＨＣＶ感染的临床诊断和发病机理的研究。

Gene fragments of E1 and E2/NS2 of HCV-Hun were cloned by RT-PCR and gene recombinant techniques from blood of HCV infected patient in Hunan province,China.In comparision cDNA sequences of our clones with those of HCV-1, HCV-J, HCV-J6and HCV-J8, gene fragments of HCV-Hun belong to genotype 1b of HCV. The fusion proteins containing the peptides encoded by HCV-E1 (MBP-HCV-E1) and HCV-E2/NS1 ( MBP-HCV-E2/NS1 ) were synthesized by E. Coli with recombinant plasmids. The specific HCV antigenicity of these fusion proteins were identified by western blotting.These fusion proteins should be useful in the study on pathogenesis and diagnosis of HCV infection.