Bcl-x基因shRNA表达质粒的构建和鉴定  

Construction and identification of ShRNA expression plasmid against human Bcl-x gene

作　　者：吴斌华[1] 陈秋生[1] 谢朝阳[1] 朱少平[1] 

机构地区：[1]广东医学院检验学院,广东湛江524000

出　　处：《现代医药卫生》2010年第15期2250-2252,共3页Journal of Modern Medicine & Health

基　　金：湛江市科技攻关计划项目(2009C3104001);广东医学院科研基金项目(XK0802)

摘　　要：目的:构建含人Bcl-x基因不同位点的一系列shRNA真核表达质粒,为以Bcl-x基因为靶点的基因治疗肿瘤研究奠定基础。方法:将人工合成的针对Bcl-x基因6个不同位点和一个混乱序列的正反链DNA序列经基因重组定向克隆插入到真核表达质粒载体pmU6中,通过酶切检测确定重组结果,并进行测序加以鉴定。结果:各对DNA序列均按正确方向插入pmU6质粒。结论:成功构建了含人Bcl-x基因6个不同位点的shRNA真核表达质粒。Objective：To construct a series of shRNA eukaryotic expression plasmids which can transcript siRNA against human Bel-x gene for future study of cancer gene therapy.Methods：7 pairs of sense and antisense sequences of siRNA which 6 pairs were a- gainst different sites of Bcl-x gene and 1 pair was scrabble DNA sequences were synthesized. They were inserted into the eukaryotic expression vector pmU6 with definite direction. They were identified by the enzyme test and sequencing test.Results ：The shRNA sequences were successfully inserted into the eukaryotic expression vector pmU6. The recombinated plasmids were identified by enzyme test and DNA sequence. Conclusion：7 siRNA transcripting plasmids against the human Bcl-x gene were constucted successfully.

关 键 词：短发夹RNA 质粒 BCL-X 

分 类 号：R446[医药卫生—诊断学]