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| 人eIF4A3真核表达载体构建及稳定表达细胞株筛选 | |
| 引用本文： | 李向云,徐祥,李秉煦,黄宏,徐云升.人eIF4A3真核表达载体构建及稳定表达细胞株筛选[J].第三军医大学学报,2011,0(18):1897-1899. |
| 作者姓名： | 李向云 徐祥 李秉煦 黄宏 徐云升 |
| 作者单位： | 1. 325000浙江温州，温州医学院附属第一医院皮肤科；400042重庆，第三军医大学大坪医院野战外科研究所一室，创伤、烧伤与复伤国家重点实验室 2. 400042重庆，第三军医大学大坪医院野战外科研究所一室，创伤、烧伤与复伤国家重点实验室 3. 温州医学院附属第一医院皮肤科,浙江温州,325000 |
| 基金项目： | 国家自然科学基金(30972800)~~ |
| 摘 要： | 目的 构建人eIF4A3重组质粒载体，筛选高表达人eIF4A3的稳定细胞株。方法RT-PCR克隆eIF4A3基因，将获得的基因片段分别连接到含有HA和c-myc标签的质粒载体上，转染HeLa细胞，经G418筛选获稳定表达细胞株，在体外翻译体系中进行体外翻译，通过免疫印迹鉴定eIF4A3的表达。结果成功构建人eIF4A3... |
| 关 键 词： | 真核起始因子4A3 稳定细胞株 体外翻译 |
Construction of human eIF4A3 eukaryotic expression plasmid and screening of stable cell line |
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| Li Xiangyun,Xu Xiang,Li Bingxu,Huang Hong,Xu Yunsheng.Construction of human eIF4A3 eukaryotic expression plasmid and screening of stable cell line[J].Acta Academiae Medicinae Militaris Tertiae,2011,0(18):1897-1899. | |
| Authors: | Li Xiangyun Xu Xiang Li Bingxu Huang Hong Xu Yunsheng |
| Institution: | Li Xiangyun1,2,Xu Xiang2,Li Bingxu1,Huang Hong2,Xu Yunsheng1(1Department of Dermatology,First Affiliated Hospital of Wenzhou Medical College,Wenzhou,Zhejiang Province,325000,2 State Key Laboratory of Trauma,Burns and Combined Injury,Department 1,Institute of Surgery Research,Daping Hospital,Third Military Medical University,Chongqing,400042,China) |
| Abstract: | Objective To construct the human eIF4A3 recombinant vector and screen the stable cell line over-expressing human eIF4A3.Methods Human eIF4A3 gene was cloned by RT-PCR.Gene fragments were linked to HA and c-myc labeled plasmid vectors to transfect HeLa cells.A stable cell line was generated by screening G418.In vitro translation was conducted in an in vitro translation system.Expression of eIF4A3 was identified by Western blotting.Results The human eIF4A3 eukaryotic expression vector was successfully constru... |
| Keywords: | eukaryotic initiation factor 4A3 stable cell line in vitro translation |
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