| 卡氏肺孢子菌主要表面糖蛋白上游保守序列基因microRNA表达载体的构建和鉴定 |
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| 引用本文： | 梅林,姚云清,朱艳玲,骆晓练,李文桂,陈雅棠.卡氏肺孢子菌主要表面糖蛋白上游保守序列基因microRNA表达载体的构建和鉴定[J].重庆医科大学学报,2010,35(2). |
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| 作者姓名： | 梅林 姚云清 朱艳玲 骆晓练 李文桂 陈雅棠 |
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| 作者单位： | 重庆医科大学附属第一医院感染科,重庆,400016;重庆医科大学附属第一医院感染科,重庆,400016;重庆医科大学附属第一医院感染科,重庆,400016;重庆医科大学附属第一医院感染科,重庆,400016;重庆医科大学附属第一医院感染科,重庆,400016;重庆医科大学附属第一医院感染科,重庆,400016 |
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| 基金项目： | 国家自然科学基金资助项目，重庆市自然科学基金资助项目 |
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| 摘 要： | 目的:构建和鉴定卡氏肺孢子菌(Pneumoeystis carinii,PC)主要表面糖蛋白(Major surface glyeoprotein,MSG)表达调控基因上游保守序列(Upstream conserved sequence,UCS)的microRNA表达载体,并在体外转染到PC中观察其对Pc的抑制作用.方法:设计并人工合成针对PC MSG-UCS基因的1对microRNA序列并定向克隆到表达载体pcDNA~(TM)6.2-GW/EmGFPmiR上,序列正确的载体用脂质体转染PC细胞,RT-PCR检测其对PC MSG-UCS基因的抑制作用,扫描电镜观察Pc的超微结构变化.结果:凝胶电泳和测序鉴定得到的产物与预期的目的基因一致.其在mRNA水平能明显抑制Pc的表达并能破坏PC的超微结构.结论:Pc MSG-UCS基因的microRNA表达载体pPC-UCSm构建成功,并能在体外抑制PC的表达.
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| 关 键 词： | 卡氏肺孢子菌 主要表面糖蛋白上游保守序列 microRNA 表达载体 |
Construction and identification of plasmid vector of microRNA on pneumocystis carinii major surface glycoprotein gene's upstream conserved sequence |
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| MEI Lin,et al.Construction and identification of plasmid vector of microRNA on pneumocystis carinii major surface glycoprotein gene's upstream conserved sequence[J].Journal of Chongqing Medical University,2010,35(2). |
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| Authors: | MEI Lin |
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| Institution: | MEI Lin,et al (Department of Infection Disease,the First Affiliated Hospital,Chongqing Medical University) |
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| Abstract: | Objective:To constructive and identify plasmid vector of microRNA on pneumocystis carinii(PC)major surface glycoprotein (MSG)gen's upstream conserved sequence(UCS),then we researched its effect after the recombinant plasmid was transfected into PC. Methods:microRNA oligonucleotides targeting PC MSG-UCS gene were chemically synthesized and inserted into plasmid vector pcDNA? 6.2-GW/EmGFPmiR after annealing.then the recombinant plasmid was transfected into PC by liposome in vitro.The PC MSG-UCS mRNA expressio... |
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| Keywords: | microRNA Pneumocystis carinii MSG-UCS MicroRNA Expression plasmid |
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