丙型肝炎病毒(HCV)非结构区NS3-5B蛋白的真核表达载体构建及其在BHK-21细胞中的表达  

Construction of the Eukaryotic Expression Vector of HCV NS3-5B and Its Expression in BHK-21 Cells

作　　者：韩佩君[1] 雷迎峰[1] 吕欣[1] 杨敬[1] 姚敏[1] 乔卿华[1] 党品香 尹文[1] 

机构地区：[1]第四军医大学基础医学部微生物学与病原生物学教研室,西安710032

出　　处：《科学技术与工程》2013年第36期10794-10797,10805,共5页Science Technology and Engineering

基　　金：国家自然科学基金项目(81171637,31172166)资助

摘　　要：通过PCR方法扩增出HCV NS3-5b全长基因序列,克隆入真核表达载体pIRES2-EGFP中,构建重组质粒pIRES2-EGFP-NS3-5b。利用脂质体将该质粒转染至BHK-21细胞,通过荧光成像和Western Blot检测NS3-5b基因的表达。结果显示成功构建了真核表达载体pIRES2-EGFP-NS3-5b,并且NS3/4A蛋白和NS5B蛋白得到特异性表达,为下一步建立HCV RdRp活性的细胞评价系统和动物模型评价系统奠定了实验和理论基础。To construct the recombinant plasmid pIRES2-EGFP-NS3-SB, HCV NS3-Sb gene was amplified by the polymerase chain reaction （PCR） and cloned into the eukaryotic expression plasmid plRES2-EGFP. Then the recombinant plasmid was transfected into BHK-21 cells by Lipofectamine 2000 and the expression of target genes were identified by fluorescence imaging and western blot analysis. The results showed that the plasmid pIRES2-EG- FP-NS3-5b was successfully constructed and NS3/4A and NSSB proteins were specifically expressed in the eukary- otic cells. It provides the basis for the establishment of a cell-based HCV RdRp evaluation system and animal mod- els development.

关 键 词：丙型肝炎病毒(hepatitis e virus HCV) NS3-5B蛋白 真核表达 

分 类 号：R373.21[医药卫生—病原生物学]