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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
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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.
作者 闫丽君 吴更
机构地区 上海交通大学生命科学技术学院
出处 《中国细胞生物学学报》 CAS CSCD 北大核心 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~~
关键词 TSC1 TSC2 稳转细胞系 真核细胞双元表达载体 Zeocin TSC 1 TSC2 stable cell line eukaryotic expression plasmid Zeocin
分类号 Q78 [生物学—分子生物学]
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中国细胞生物学学报
2012年 第12期

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