稳定表达TSC1/TSC2蛋白质复合物的293T细胞系的建立  

Generation of Stable Production of TSC1/TSC2 Protein Complex from 293T Cell Line

作　　者：闫丽君[1] 吴更[1] 

机构地区：[1]上海交通大学生命科学技术学院,微生物代谢国家重点实验室,上海200240

出　　处：《中国细胞生物学学报》2012年第12期1215-1219,共5页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(No.30821005)资助项目~~

摘　　要：我们使用Clonetech的同源重组酶连接人TSC1、TSC2全长蛋白编码cDNA ORF到pBudCE4.1真核细胞双元表达载体上,用脂质体Lipofectamine 2000介导重组质粒pBudCE4.1/TSC2/TSC1导入293T细胞,用含125μg/mL Zeocin的培养基筛选稳定表达TSC1/TSC2蛋白的细胞株,并用Western blot方法鉴定稳转细胞株的稳定性。该实验成功建立了稳定表达TSC1/TSC2蛋白的293T细胞系,从而为今后研究TSC1/TSC2蛋白的结构与功能提供实验基础。The cDNA ORFs of human TSC1 and TSC2 were cloned into the two multi-cloning sites of the eukaryotic expression plasmid pBudCE4.1 vector by In-Fusion~ HD Cloning Kit. The recombinant vector was transfected into 293T cells by Lipofectamine 2000, under the pressure of Zeocin. Stable clones were picked and an- alyzed by Western blot. In this study, stable production of TSC1/TSC2 protein complex from the 293T cell line was established, which would be a great help for further structural and functional research of the TSC 1/TSC2 complex.

关 键 词：TSC1 TSC2 稳转细胞系 真核细胞双元表达载体 Zeocin 

分 类 号：Q78[生物学—分子生物学]