摘要
目的构建丙型肝炎病毒5′非翻译区（HCV 5′UTR）调控绿色荧光蛋白真核表达质粒，并在HepG2细胞中表达。方法通过PCR扩增，获得HCV基因组HCV 5′UTR完整序列及C区序列的部分基因片断。将此片断插入pEGFP-NI载体多克隆位点区，构建受HCV 5′UTR调控的绿色荧光蛋白真核表达质粒．应用脂质体转染技术转染HepG2细胞并观察荧光。结果成功构建受HCV 5′UTR调控的绿色荧光蛋白真核表达质粒，并在HepG2细胞中观察到绿色荧光蛋白的表达。结论该载体的成功构建，可以用于直观地评价针对HCV 5′UTR的基因药物的效果。
Objective To establish an in vitro screening system for testing effects of genetic drugs targeting at 5′ UTR of HCV genome. Methods In order to construct the plasmid expressing HCV-gfp fusion RNA, the target gene fragment was obtained by PCR amplification and inserted into pEGFP-N1 reporter vector. The structure of constructed plasmid was confirmed by electrophoresis analysis and DNA sequencing. The function of construct was confirmed by lipofectamine-mediated transient expression in HepG2 cells. Results DNA sequencing showed that the inserted fragment of the constructed plasmid was the same as the template HCV genome. The HepG2 cells transfected with the constructed plasmid could express reporter gene of gfp. Conclusions The results showed that the author successfully constructed the plasmid expressing gfp gene controlled by HCV 5′ UTR and established an in vitro testing system for evaluating HCV 5′ UTR specific nucleic acid drugs.
出处
《世界感染杂志》
2006年第1期16-18,共3页
World Journal of Infection
基金
国家自然科学基金（30400374）
