| 稳定表达TSC1／TSC2蛋白质复合物的293T细胞系的建立 |
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| 引用本文： | 闫丽君,吴更. 稳定表达TSC1／TSC2蛋白质复合物的293T细胞系的建立[J]. 细胞生物学杂志, 2012, 0(12): 1215-1219 |
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| 作者姓名： | 闫丽君 吴更 |
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| 作者单位： | 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 |
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| 基金项目： | 国家自然科学基金（No.30821005）资助项目 |
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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eDNA0RF到pBudCE4．1真核细胞双元表达载体上，用脂质体Lipofectamine2000介导重组质粒pBudCE4．1／TSC2／TSC1导入293T细胞，用含125μg／mLzeocin的培养基筛选稳定表达TSC1/TSC2蛋白的细胞株，并用Westemblot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。
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| 关 键 词： | TSC1 TSC2 稳转细胞系 真核细胞双元表达载体 Zeocin |
Generation of Stable Production of TSC1/TSC2 Protein Complex from 293T Cell Line |
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| Yan Lijun,Wu Geng. Generation of Stable Production of TSC1/TSC2 Protein Complex from 293T Cell Line[J]. Chinese Journal of Cell Biology, 2012, 0(12): 1215-1219 |
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| Authors: | Yan Lijun Wu Geng |
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| Affiliation: | (School of Life Sciences ＆ Biotechnology, State Key Laboratory of Microbial Metabolism Shanghai Jiao Tong University, Shanghai 200240, China) |
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| Abstract: | 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. |
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| Keywords: | TSC 1 TSC2 stable cell line eukaryotic expression plasmid Zeocin |
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