持续表达HCV NS5B的Bel-7402细胞系的构建与鉴定 Establishment of the Bel-7402 Lines Constitutively Expressing Enzymatically Active HCV NS5B.pdf

第31卷第4期 江西农业大学学报 V01．31，No．4 Acta Universitatis 2009年8月 Agricuhurae Jiangxiensis Aug．，2009 文章编号：1000—2286(2009)04—0717—06 NS5 持续表达HCVB的Bel一7402 细胞系的构建与鉴定 孔令保，吴晓玉，李昆太，徐 波，刘好桔，熊舒琚 (江西农业大学生物科学与工程学院，江西南昌330045) 摘要：HCV G418筛选转染细胞。阳性细胞经基因组PCR、RT—PCR和WesternBlot鉴定，证实获得持续表达NS5B的Bel 依赖的RNA聚合酶活性。持续表达有聚合酶活性的HCVNS5B的Bel一7402细胞系的建立为进一步研究 Ns5B催化的HCV复制机制和NS5B抑制剂的筛选奠定基础。 关键词：HCV；NS5B；Bel一7402；持续表达；活性 中图分类号：Q785；R512．6+3文献标识码：A oftheBel··—7402 Establishment Lines Constitutively ActiveHCVNS5B ExpressingEnzymatically KONG Ling—bao，WUXiao—yu，LIKun—tai， XU Bo，LIU Hao-ju，XIONGShu—jan ofLifeScienceand (College Engineering，JAU，Nanchang330045，China) Abstract：HCV RNA B)is forviral and RNA—dependentpolymerase(NS5strictlyrequired replication allattractiveforantiviral of celllines represents target drugdevelopment．Establishmenthepatic constitutively HCVNS5Bshouldbe for the expressing ac6ve helpful mechanismsof enzynmtically studying vlra／replication in tensand ofantiviral forthe recombinantvector hepatic development drugs．The pcDNA一5B expressing HCVNS5B was withtherestriction theScaI—linearized