E2F1基因真核表达载体的构建及其对CD2AP启动子的影响  

Construction of an eukaryotic vector expressing E2F1 gene and its effect on CD2AP promoter

作　　者：邹黎[1] 金蕊[1] 任伟[1] 王怡[1] 朱亮华[1] 李悦[1] 周国平[1] 

机构地区：[1]南京医科大学第一附属医院儿科,210036

出　　处：《江苏医药》2011年第2期129-131,共3页Jiangsu Medical Journal

基　　金：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)

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。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 sequence analysis and Western blot indicated that an eukaryotic vector expressing E2F1 gene was successfully constructed . Over-expression of E2F1 increased the promoter activity of CD2AP.Conclusion E2F1 is correctly expressed in HEK-293 cells. E2F1 transcription factor can enhance human CD2AP promoter activity.

关 键 词：E2F1 CD2相关蛋白 肾小球硬化 

分 类 号：R729[医药卫生—儿科]