| E2F1基因真核表达载体的构建及其对CD2 AP启动子的影响 |
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| 引用本文： | 邹黎,金蕊,任伟,王怡,朱亮华,李悦,周国平.E2F1基因真核表达载体的构建及其对CD2 AP启动子的影响[J].江苏医药,2011,37(2). |
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| 作者姓名： | 邹黎 金蕊 任伟 王怡 朱亮华 李悦 周国平 |
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| 作者单位： | 南京医科大学第一附属医院儿科,210036 |
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| 基金项目： | 国家自然科学基金资助项目(30872804,30570863); 江苏省自然科学基金资助项目(BK2007244); 江苏省“科教兴卫工程”医学重点人才项目资助(RC2007050) |
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| 摘 要： | 目的构建E2F1基因真核表达质粒,并初步探讨E2F1对CD2相关蛋白(CD2AP)启动子的作用。方法 RT-PCR扩增E2F1基因,构建含E2F1基因的重组真核表达质粒,重组质粒转染人胚肾HEK-293细胞。Western blot检测E2F1蛋白表达;双荧光素酶报告基因检测E2F1过表达后人CD2AP启动子的活性。结果核酸序列分析及Western blot结果证实,成功构建含E2F1基因的重组真核表达质粒;过表达E2F1能增强CD2AP启动子的活性。结论 E2F1编码基因在HEK-293细胞中获得正确表达,且E2F1蛋白可促进人CD2AP启动子的转录活性。
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| 关 键 词： | E2F1 CD2相关蛋白 肾小球硬化 |
Construction of an eukaryotic vector expressing E2F1 gene and its effect on CD2AP promoter |
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| ZOU Li,JIN Rui,REN Wei,et al..Construction of an eukaryotic vector expressing E2F1 gene and its effect on CD2AP promoter[J].Jiangsu Medical Journal,2011,37(2). |
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| Authors: | ZOU Li JIN Rui REN Wei |
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| Institution: | ZOU Li,JIN Rui,REN Wei,et al.Department of Pediatrics,First Affiliated Hospital,Nanjing Medical University,Nanjing 210029,CHINA |
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| Abstract: | Objective To construct an eukaryotic expression vector of E2F1 and investigate the effect of E2F1 on the promoter activity of CD2AP.Methods E2F1 gene was amplified by RT-PCR.Recombinant eukaryotic expression plasmids encoding E2F1 were constructed.HEK-293 cells were transfected with the recombinant plasmids and Western blot was performed to evaluate the protein expression of E2F1.The human CD2AP promoter activity after over-expressing E2F1 was examined by dual luciferase reporter gene analysis.Results DNA s... |
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| Keywords: | E2F1 CD2-associated protein Glomerularsclerosis |
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