用Gateway系统构建携带Grb2-SH2基因重组慢病毒载体  

Construction of Grb2-SH2 genetic restructuring-slow virus carrier with Gateway system

作　　者：蔡忠良[1] 阴继凯[1] 韩腾龙[2] 张健[2] 鲁建国[1] 

机构地区：[1]第四军医大学附属唐都医院普通外科,陕西西安710038 [2]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710032

出　　处：《现代生物医学进展》2012年第4期616-618,共3页Progress in Modern Biomedicine

基　　金：国家自然青年科学基金资助项目(30901457)

摘　　要：目的:构建携带Grb2-SH2基因重组慢病毒载体。方法:把Grb2-SH2基因从合成的工程载体PUC-57中导到入门载体pentr-3C中,再利用LR反应重组到目的载体plenti,通过酶切和测序分析对比验证Grb2-SH2基因后,将plenti-Grb2-SH2质粒利用慢病毒转染系统转染人胚胎肾上皮细胞株293T细胞获得携带Grb2-SH2慢病毒载体。结果:构建的重组质粒经酶切及测序和分析比对鉴定正确,目的基因片段大小约为500bp;该质粒与包装质粒共转染293T细胞获取的慢病毒,转染效率约为90%。结论:成功构建转载质粒plenti-Grb2-SH2及携带Grb2-SH2基因慢病毒载体。Objective： Construct Grb2-SH2 genetic recombination slow virus carrier.Methods： Put the Grb2-SH2 genes into introductory carrier pentr-3 C from synthetic engineering carrier PUC-57,reuse LR reaction to recombinate into the purpose plenti carrier,after comparing and testing gene Grb2-SH2 through the enzyme cut and sequencing analysis and comparison,use slow virus carrying system to transfect human embryonic renal epithelial cell lines 293 T cells acquring Grb2-SH2 slow virus carrier.Results： The restructuring plasmid proved to be correct by the enzyme cut and sequencing analysis and the purpose genetic fragments is about 500 bp;The efficiency of the plasmid and packaging plasmid transfect 293 T cells get slow is about 90%.Conclusion： Successful construction reproduced plasmid plenti-Grb2-SH2 and Grb2-SH2 gene slow virus carrier.

关 键 词：生长因子受体连接蛋白 重组慢病毒载体 表皮生长因子受体 

分 类 号：Q75[生物学—分子生物学] Q78