肝特异性表达HCV5′NCR调控荧光素酶质粒的构建及表达

Construction and Expression of Liver-Specific Expression Plasmid of Luciferase Controlled by HCV5′NCR Region

小鼠白蛋白是肝组织特异性表达的蛋白 ,这种特异性是由白蛋白启动子所介导的 .以2 2 35A- 1质粒为模板 ,通过 PCR扩增获得小鼠白蛋白启动子 /增强子基因片段 ,用小鼠白蛋白启动子 /增强子基因片段取代 p HCV- neo4质粒 (含 HCV5′NCR调控荧光素酶基因 )的 CMV启动子 ,构建了一种白蛋白启动子启动转录的 HCV5′NCR调控荧光素酶表达质粒 (p A1 b- HCV) .该质粒能在小鼠肝癌细胞中表达且较小鼠其它癌细胞中表达水平明显增高 ,表明成功地构建了肝特异性表达的 HCV5′NCR调控荧光素酶表达质粒 .该研究为建立肝特异性表达的 HCV5′NCR转基因小鼠模型奠定了基础 ,对评价

Mouse albumin is a kind of liver tissue specific expression protein which is mediated by albumin promoter.Mouse albumin promoter/enhancer gene was obtained from the plasmid 2235A 1 by PCR and substituted the CMV promoter of plasmid pHCV neo4 which contained luciferase gene controlled by HCV5′NCR using standard protocols.The plasmid(pA1b HCV) expressing luciferase controlled by HCV5′NCR,which was initiated by albumin promoter/enhancer,was constructed.This plasmid could express luciferase activity in Hepal 6 cells (mouse,hepatoma) and showed higher levels of expression than that in other cells such as LA795 and B16 cells.These results suggest that the liver specific expression plasmid expressing luciferase controlled by HCV5′NCR was successfully constructed.This has provided a basis to establishing the HCV5′NCR transgene mouse model of liver specific expression.It is of great significance to evaluate the antisense drugs against HCV and liver targeted delivery systems.