丙型肝炎病毒结构蛋白在痘苗病毒中的表达

Expression of hepatitis C viral structure protein by recombinant vaccinia virus

为研究中国丙型肝炎病毒（ＨＣＶ）的抗原性及在细胞内的加工，将丙型肝炎病毒（ＨＣＶ）５’非编码区（ＮＴＲ）和结构基因（Ｃｏｒｅ＋Ｅ１＋Ｅ２／ＮＳ１）插入痘苗病毒表达载体ｐＪＳＡ１１７５中，转染ＴＫ－１４３细胞，经纯化得到丙型肝炎（ＨＣＶ）重组痘苗病毒ｖＪＳＡ１１７５ＣＥ株。Ｓｏｕｔｈｅｒｎｂｌｏｔ杂交表明，ＨＣＶ结构基因存在于痘苗病毒之中。Ｗｅｓｔｅｒｎｂｌｏｔ分析发现，ｖＪＳＡ１１７５ＣＥ表达蛋白带位于９０ｋＤａ，为一多聚蛋白；此蛋白为分泌型。

The hepatitis C viral (HCV) DNA fragment coding for the core and two glycoproteins E1+E2/NS1 were inserted into vaccinia virus vector pJSA1175, under the control of the promoter 7.5K. 143TK- cells transfected with the HCV recombinant plasmid pJSA1175CE in the presence of infectious TK+ vaccinia virus (Tiantan stain) yielded recombinant vaccinia virus and expressed a ～90kDa polyprotein. Southern blot hybridization showed that the inserted HCV cDNA was located at the vaccinia virus TK+ gene, J fragment. One polypeptide band between 106 and 80kDa was revealed in SDS-PAGE which reacted with anticore murine serum. The polypeptide could be secreted into the medium, the total amount was about the same as in the cells.