风疹病毒JR23株E1包膜糖蛋白基因的表达分析

摘要

To construct the eukaryot ic expression system of E1 glycopr otein of rubella virus(RV)JR23 str ain and to study the biological an d immunological properties of the expressed product,the E1 gene was amplified by RT-PCR,sequenced and recombinated with the pBluescript ⅡSK+ vectorThe recombinated vecto r was co-transfected into BHK21 ce ll with vaccinia virus which carri es the gene of T7 RNA polymeraseT he expressed protein was characteri zed with hemadsorption,immunohisto chemistry and indirect immunofluor escence stainingThe results showe d that the recombinated vector,pBS K+-RV E1,was proved containing E1 gene by enzyme digestion and seque ncingThe expressed protein could be detected both in cytoplasm amd on membrane of the transfected cel ls by hemadsorption,immunohistoche mistry and indirect immunofluoresc enceThe E1 protein mainly focused in cytoplasm near the nucleusThes e data proved that the expression vector,pBSK+-RV E1,was successfull y constructed and the glycoprotein E1 of RV JR23 strain was effective ly expressed in BHK21 cell culture This study provides foundations f or studies on the relationships be tween structure and function of RV gene,between structure and functio n of RV proteins,and on the subuni t vaccine of rubella