一种四环素严紧控制的真核表达系统  

An eucaryotic expression system tightly controlled by tetracycline

作　　者：铁茹[1] 陈宝莹[2] 曲萍[1] 戚朦[1] 杨东海[1] 于军[1] 

机构地区：[1]第四军医大学基础部基础医学教学实验中心,西安710032 [2]第四军医大学唐都医院放射科

出　　处：《山西医科大学学报》2011年第5期361-365,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30801385);陕西省科技攻关课题(2009K12-01)

摘　　要：目的构建一种严紧调控的表达系统,用于在真核细胞内表达外源性兴趣基因。方法融合四环素诱导元件、人工细菌染色体(BAC)以及Gateway技术,建立了一种新的表达体系。并且将该体系进行了测试。结果兴趣基因编码的β-catenin-ERα融合蛋白经荧光素酶活性检测和Western杂交分析,其表达被四环素或强力霉素严紧控制,在表达载体pE11-IGR-β-catenin-ERα中,EGFP的表达量恒定,可以作为流式细胞术有效的筛选标记。结论该系统可以作为一种有效的工具,用于在真核细胞中表达外源性兴趣基因,并且实现严紧的表达调控。Objective To construct a stringently controlled expression system for exogenous gene of interest in eukaryotic cells. Methods A new expression system was established by fusing tetracycline-inducible elements,bacterial artificial chromosome（BAC）and Gateway technology and then tested. Results The fusion protein β-catenin-ERα encoded by gene of interest was tested by luciferase assay and Western blot,and its expression was tightly controlled by tetracycline or deoxycycline.In the final expression vector BAC E11-IGR-β-catenin-ERα,the expression of EGFP was stable,which could be an effective marker for FACS selection. Conclusion This system could be used as an efficient tool for the expression of exogenous gene of interest in eukaryotic cells in a stringently-controlled manner.

关 键 词：细菌人工染色体 四环素 强力霉素 

分 类 号：Q78[生物学—分子生物学]