卡氏肺孢子胸腺嘧啶核苷酸合酶基因siRNA表达载体的构建和鉴定  

Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii thymidylate synthase gene

作　　者：唐滢浍[1] 姚云清[1] 谭燕[1] 王莉妮[1] 李文桂[1] 刘成伟[1] 陈雅棠[1] 

机构地区：[1]重庆医科大学附属第一医院感染科,重庆400016

出　　处：《重庆医科大学学报》2009年第7期877-879,共3页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(30471512);重庆市医学科技计划项目(04-2-100)

摘　　要：目的:构建和鉴定大鼠卡氏肺孢子(Pneumocystis carinii,PC)胸腺嘧啶核苷酸合酶(Thymidylate synthase,TS)基因的siRNA(Short interfering RNA)表达载体。方法:人工合成针对PCTS基因的1对shRNA(Short hairpin RNA)序列并定向克隆到空载体pTZU6+1上,构建siRNA表达重组质粒pPC-TS,并采用双酶切产物凝胶电泳和基因测序法鉴定。结果:酶切电泳和基因测序鉴定得到的产物与预期的目的基因一致。结论:成功构建卡氏肺孢子胸腺嘧啶核苷酸合酶基因siRNA表达载体。Objective：To construct and identify a plasmid vector of short interfering RNA （siRNA） on pneumocystis carinii （PC） thymidylate synthase. Methods： Short hairpin RNA oligonucleotides of thymidylate synthase were chemically synthesized and inserted into plasmid vector pTZU6＋ 1 after annealing. The recombinant plasmid, pPC-TS, transformed into E. coil. TOP10 and amplified, was digested by restriction endonucleases Hind Ⅲ and EcoRⅠ and identified by gel electrophoresis and DNA sequencing. Results： Gel electrophoresis and DNA sequencing showed that the recombinant plasmid containing the correct and full shRNA oligonucleotide. Conclusion ： The siRNA plasmid, pPC-TS, was constructed successfully.

关 键 词：卡氏肺孢子 胸腺嘧啶核苷酸合酶基因 SIRNA表达载体 

分 类 号：R730.23[医药卫生—肿瘤]