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| 顶体素结合蛋白真核表达载体的构建及其在肝癌细胞株的表达 | |
| 引用本文： | 罗彬,云翔,林永达,肖绍文,闫光华,何少健,贺菽嘉,陈芳,谢小薰.顶体素结合蛋白真核表达载体的构建及其在肝癌细胞株的表达[J].细胞与分子免疫学杂志,2011,27(10):1072-1074. |
| 作者姓名： | 罗彬 云翔 林永达 肖绍文 闫光华 何少健 贺菽嘉 陈芳 谢小薰 |
| 作者单位： | 1. 广西医科大学组织胚胎学教研室,广西南宁,530021 2. 广西医科大学第一附属医院,广西南宁,530021 3. 广西医科大学生化教研室,广西南宁,530021 |
| 基金项目： | 国家自然科学基金资助项目(30760055,81060207); 广西高发疾病研究创新性团队基金资助项目(桂教人[2007-71]号); 广西自然科学基金(桂科自0832144); 广西教育厅立项项目(201010LX029),广西教育厅研究生创新计划项目(2009105981001M190); 广西医学科学实验中心开放项目(KFJJ2010-33); 广西大型仪器网协助项目(6912008104) |
| 摘 要： | 目的:构建ACRBP真棱表达载体,建立稳定表达ACRBP的人肝癌细胞株,为后继研究该基因的功能奠定基础.方法:以pMAL-C2/ACRBP重组质粒作为模板进行PCR,扩增出ACRBP编码区cDNA,并与真核表达载体pEGFP-N1连接,构建pEGFP-N1/ACRBP重组质粒.通过Fugene HD将该重组质粒转染至ACRBP表达阴性的肝癌细胞株HepG2,经G418筛选阳性克隆获得稳定转染株.RT-PCR和免疫组织化学方法检测HepC-2细胞中ACRBP的表达情况.结果:pEGFP-N1/ACRBP真核表达载体经测序证实插入序列完全正确;ACRBP基因已经稳定转染至HepG2细胞中并获得表达.结论:成功构建了pEGFP-N1/ACRBP真核表达载体并将其转染HepG2后,能稳定地表达ACRBP. |
| 关 键 词： | ACRBP 基用克隆 转染 |
Construction of eukaryotic expression vector encoding ACRBP and its expression in hepatocarcinoma cells |
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| Luo Bin,Yun Xiang,Lin Yong-da,Xiao Shao-wen,Yan Guang-hua,He Shao-jian,He Shu-jia,Chen Fang,Xie Xiao-xun.Construction of eukaryotic expression vector encoding ACRBP and its expression in hepatocarcinoma cells[J].Journal of Cellular and Molecular Immunology,2011,27(10):1072-1074. | |
| Authors: | Luo Bin Yun Xiang Lin Yong-da Xiao Shao-wen Yan Guang-hua He Shao-jian He Shu-jia Chen Fang Xie Xiao-xun |
| Institution: | Department of Histology and Embryology, Guangxi Medical University, Nanning, China. |
| Abstract: | AIM: To construct the eukaryotic expression vector pEGFP-N1/ACRBP and stably express ACRBP in human hepatocarcinoma cells, providing functional clues for ACRBP. METHODS: A recombinant plasmid pMAL-C2/ACRBP was used as a template to amplify ACRBP cDNA. The PCR product was ligated into an eukaryotic expression vector pEGFP-N1 to construct a recombinant plasmid pEGFP-N1/ACRBP. Then the pEGFP-N1/ACRBP was transfected by Fugene HD into ACRBP-negative HepG2 cells. The stably transfected clones were selected by G4... |
| Keywords: | ACRBP gene cloning transfection |
| 本文献已被 CNKI 万方数据 PubMed 等数据库收录！ | |