四环素调控的真核表达体系Hep3B Tet-On细胞株的建立  

Establishment of Hep3B cell line controlled by the Tet-On regulatory system

作　　者：王俊茹[1] 覃文新[1] 舒惠群[1] 潘勇[1] 张玉燕[1] 万大方[1] 

机构地区：[1]上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海20032

出　　处：《肿瘤》2002年第3期191-193,共3页Tumor

基　　金：国家"8 6 3"生物高技术资助项目 (Z1 9 0 2 0 1 0 1 );国家重点基础研究发展规划" 973"资助项目(G 1 9980 51 2 0 9)

摘　　要：目的　建立可受四环素诱导调控的高表达外源基因的真核表达体系Hep3BTet On细胞株 ,用于基因功能的研究。方法　将pTet On质粒用脂质体介导法转染Hep3B细胞 ,经G4 18筛选后的克隆用有限稀释法进行单克隆化。单克隆分别扩增后 ,瞬时转染pTRE luc(编码荧光素酶蛋白 )质粒 ,Dox诱导表达后 ,检测荧光素酶活性 ,逐一筛选四环素调控高表达外源基因的Hep3BTet On细胞株。结果　成功构建了一株受Dox调控的高表达低背景的Hep3BTet On细胞株。结论　Hep3BTet On细胞株可用于外源基因的真核调控高表达 ,为研究真核基因功能提供了一种有力的实验手段。Objective To establish tetracycline controlled inducible system (Tet On) in Hep3B cell. Methods The Hep3B cells were transfected with pTet On vector by using liposome transfection reagent. After 48 hours, the transfected cells were selected in medium containing G418. 20 days later, G418 resistant clones were isolated. All individual G418 resistant clones were screened by transient transfection with pTRE luc for clones with low background and high induction of luciferase in response to Dox. Results One Hep3B cell line, which exhibited high levels of induction and high gene expression levels, was obtained. Conclusion The Hep3B cell line can be used to highly express eukaryotic gene and this Tet On system is available for use in eukaryotic gene function studies.

关 键 词：真核表达体系 Hep3BTet-On细胞株 诱导表达 四环素调控 基因治疗 基础研究 基因表达 

分 类 号：Q786[生物学—分子生物学]