编码Sema3a基因慢病毒载体的构建  

Construction of lentiviral vectors carrying Sema3a gene

作　　者：陈仁华[1,2] 李毅刚[2] 王祥贵[1] 许祖芳[1] 陈友佳[1] 李威[2] 

机构地区：[1]江西省赣州市人民医院心内科,赣州341000 [2]上海交通大学附属新华医院心内科,上海200092

出　　处：《江西医药》2012年第11期956-958,共3页Jiangxi Medical Journal

摘　　要：目的构建Sema3a基因慢病毒表达载体,为转染原代心肌细胞及进一步研究奠定基础。方法 NCBI-Gene数据库检索Sema3a基因,设计合成PCR反应需要的上下游引物并扩增。扩增产物通过凝胶电泳并回收,SwaI酶切载体行酶切后连接,CaCl2法制备感受态大肠杆菌,转化扩增后,挑选克隆进行重组鉴定。结果PCR扩增和测序结果证实成功构建Sema3a的慢病毒载体检测并包装慢病毒。结论成功建立Sema3a重组慢病毒载体的三质粒包装系统。Objective To construct lentiviral vectors carrying Sema3a gene.Methods Retrieved Sema3a gene sequendes from the NCBI-Gene bank, designed and synthesized the upstream and downstream primers PCR needed. Digested and connected amplification products recycled by gel electrophoresis. After conversed and amplified competent E. coli prepared through CaC12, the selection of clone were reorganized and identified.Results PCR amplification and sequencing results confirmed successful construction of a lentiviral vector detection in Sema3a and packaging lentivirus.Conelusion The successful establishment of three plasmids Sema3a recombinant lentiviral vector packaging systems.

关 键 词：SEMA3A SwaI酶 慢病毒裁体 

分 类 号：R-33[医药卫生]