卡氏肺孢子表面糖蛋白siRNA表达载体的构建和鉴定  

Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface gene

作　　者：王莉妮[1] 姚云清[1] 谭燕[1] 唐滢浍[1] 李文桂[1] 马良[1] 陈雅堂[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》2008年第5期517-519,525,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金(30471512)

摘　　要：目的:构建和鉴定卡氏肺孢子(Pneumocystis carinii,PC)主要表面糖蛋白(Major surfce glycoprotein,MSG)基因短干扰性RNA(short interfering RNA,siRNA)表达载体。方法:设计并人工合成针对PC MSG基因的短发夹状(Short hairpin RNA,shRNA)序列并定向克隆到siRNA表达载体pTZU6+1上,采用双酶切后凝胶电泳与DNA测序法进行鉴定。结果:酶切和测序鉴定得到的产物与预期目的基因一致。结论:成功构建卡氏肺孢子主要表面糖蛋白基因siRNA表达载体。Objective：To construct and identify a plasmid vector of short interfering RNA（siRNA） on pneumocystis carinii（PC） major surface glycoprotein（MSG） gene. Methods：Short hairpin RNA（shRNA） oligonucleotide targeting PC MSG gene which was chemically synthesized and inserted into RNAi-Ready plasmid vector pTZU6＋1 after annealing.the recombinant plasmid, pPC-MSG,transgormed into E.coil.TOP10 and amplified,was digested by restriction endonucleases Hind Ⅲ and EcoR Ⅰ and identified by gelelectrophoresis and DNA sequencing. Results：Gel electrophoresis and DNA sequencing showed that the recomhinant plasmid containing the correct and full shRNA oligonucleotide. Conclusion：The siRNA plasmid,pPC-MSG was constructed successfully.

关 键 词：卡氏肺孢子 主要表面球蛋白 SIRNA 表达载体 

分 类 号：R531.9[医药卫生—内科学]