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-PC...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+ vectorThe recombinated vecto r was co-transfected into BHK21 ce ll with vaccinia virus which carri es the gene of T7 RNA polymeraseT he expressed protein was characteri zed with hemadsorption,immunohisto chemistry and indirect immunofluor escence stainingThe results showe d that the recombinated vector,pBS K+-RV E1,was proved containing E1 gene by enzyme digestion and seque ncingThe expressed protein could be detected both in cytoplasm amd on membrane of the transfected cel ls by hemadsorption,immunohistoche mistry and indirect immunofluoresc enceThe E1 protein mainly focused in cytoplasm near the nucleusThes 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展开更多
文摘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+ vectorThe recombinated vecto r was co-transfected into BHK21 ce ll with vaccinia virus which carri es the gene of T7 RNA polymeraseT he expressed protein was characteri zed with hemadsorption,immunohisto chemistry and indirect immunofluor escence stainingThe results showe d that the recombinated vector,pBS K+-RV E1,was proved containing E1 gene by enzyme digestion and seque ncingThe expressed protein could be detected both in cytoplasm amd on membrane of the transfected cel ls by hemadsorption,immunohistoche mistry and indirect immunofluoresc enceThe E1 protein mainly focused in cytoplasm near the nucleusThes 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
