| JTV1基因真核表达质粒的构建及其在K562细胞中的表达 |
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| 引用本文： | 吴燕,刘北忠,王翀,钟梁,朱丹,王春光,金丹婷,高艳军,黎亮.JTV1基因真核表达质粒的构建及其在K562细胞中的表达[J].粉末涂料与涂装,2011,24(1). |
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| 作者姓名： | 吴燕 刘北忠 王翀 钟梁 朱丹 王春光 金丹婷 高艳军 黎亮 |
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| 作者单位： | 重庆医科大学医学检验系临床检验诊断学教育部重点实验室; |
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| 基金项目： | 国家自然科学基金(30300449); 国家中医药管理局(02-03ZP52); 重庆医科大学课题(XBYB2007104) |
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| 摘 要： | 目的 构建JTV1基因真核表达质粒并稳定转染人白血病细胞系K562,检测转染细胞中JTV1基因mRNA和蛋白的表达水平及其对K562细胞增殖的影响。方法从人外周血单个核细胞中克隆JTV1基因,并将其插入pcDNA3.1表达载体中,构建真核重组表达质粒pcDNA3.1-JTV1,经脂质体介导转染K562细胞,采用RT-PCR和Western blot法鉴定转染细胞中JTV1基因mRNA和蛋白的表达水平;MTT法检测JTV1稳定表达对K562细胞增殖的影响。结果重组表达质粒pcDNA3.1-JTV1经双酶切及测序证实,目的基因已插入质粒中;人JTV1基因能在K562细胞中稳定表达;JTV1具有抑制K562细胞增殖的作用。结论已成功构建了JTV1基因真核表达质粒,并获得了稳定表达人JTV1基因的K562细胞克隆,为进一步研究人JTV1基因的功能及其与白血病细胞增殖及凋亡的相关性提供了细胞模型。
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| 关 键 词： | JTV1基因 真核细胞 基因表达 K562细胞 细胞增殖 |
Construction of Eukaryotic Expression Vector for JTV1 Gene and Its Expression in K562 Cells |
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| WU Yan,LIU Bei-zhong,WANG Chong,ZHONG Liang,ZHU Dan,WANG Chun-guang,JIN Dan-ting,GAO Yan-jun,LI Liang.Construction of Eukaryotic Expression Vector for JTV1 Gene and Its Expression in K562 Cells[J].Chinese Journal of Biologicals,2011,24(1). |
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| Authors: | WU Yan LIU Bei-zhong WANG Chong ZHONG Liang ZHU Dan WANG Chun-guang JIN Dan-ting GAO Yan-jun LI Liang |
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| Affiliation: | WU Yan,LIU Bei-zhong,WANG Chong,ZHONG Liang,ZHU Dan,WANG Chun-guang,JIN Dan-ting,GAOYan-jun,LI Liang(Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education,Faculty of Laboratory Medicine in Chongqing Medical University,Chongqing 400016,China) |
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| Abstract: | Objective To construct a eukaryotic expression vector for JTV1 gene,stably transfect to human leukemia cellline K562,determine the expression levels of JTV1 mRNA and protein and investigate the effect of expressed product on proliferationof K562 cells. Methods JTV1 gene was cloned from human peripheral blood mononuclear cells(PBMCs)and inserted into expression vector pcDNA3. 1. The constructed plasmid pcDNA3. 1-JTV1 was transfected to K562 cells in mediation of liposome. The ex-pression levels of JTV1 mRNA ... |
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| Keywords: | JTV1 gene Eukaryocytes Gene expression K562 cells Cell proliferation |
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