Tet-on和Cre/loxP系统双重调控HCV NS5B真核表达载体的设计与构建  

Design and construction of HCV NS5B eukaryotic expression vector with Tet-on and Cre/loxP system

作　　者：马玉[1] 赵亚[1] 孙梦宁[1] 兰海云[1] 吕欣[1] 杨敬[1] 雷迎峰[1] 姚敏[1] 尹文[2] 汪爱勤[1] 

机构地区：[1]第四军医大学基础部微生物学与病原生物学教研室 [2]第四军医大学中心实验室,陕西西安710032

出　　处：《现代生物医学进展》2010年第19期3606-3608,共3页Progress in Modern Biomedicine

基　　金：国家高技术研究发展计划专题(863计划专题)(2007AA02Z154);国家自然科学基金(30872218);陕西省科技攻关计划(2010K15-03-04)

摘　　要：目的:构建可受Tet-on和Cre/loxP系统双调控的HCV NS5B真核表达载体,为建立可严格调控HCVNS5B蛋白表达的转基因小鼠奠定基础。方法:以真核表达载体pBI-3为载体构建骨架,在其启动子下游依次插入luc报告基因、BGHpA和NS5B基因片段,并分别在luc报告基因上游和BGHpA尾下游引入一个loxP位点。结果:成功构建了可受Tet-on和Cre/loxP系统双调控的HCV NS5B真核表达载体pBI-3/luc-BGHpA-NS5B。结论:pBI-3/luc-BGHpA-NS5B真核表达载体的成功构建为可严格调控HCV NS5B蛋白表达转基因小鼠的建立打下了良好的基础。Objective： To construct a HCV NS5B eukaryotic expression vector dually-controlled by Tet-on and Cre/loxP for producing transgenic mice which can strictly regulate expression of HCV NS5B. Methods： By utilizing the eukaryotic expression vector pBI-3 as vector framework, the luc reporting, the BGH pA tail and the NS5B fragment were orderly inserted in the downstream of the promoter, and a loxP site was introduced respectively in the upstream of the luc reporting and downstream of the BGH pA tail. Results： The HCV NS5B eukaryotic expression vector pBI-3/ luc-BGH pA-NS5B, which could be dually-controlled by Tet-on and Cre/loxP, were successfully constructed. Conclusion： The establish of the pBI-3/ luc-BGH pA-NS5B provides the basis for the establishment of transgenic mice that could strictly regulate the expression of HCV NS5B.

关 键 词：HCV NS5B 转基因小鼠 TET-ON Cre/loxP 

分 类 号：Q78[生物学—分子生物学] Q503