Construction and identification of plasmid vector of microRNA on pneumocystis carinii major surface glycoprotein gene’s upstream conserved sequence

MEI Lin; et al

doi:10.13406/j.cnki.cyxb.2010.02.007

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Construction and identification of plasmid vector of microRNA on pneumocystis carinii major surface glycoprotein gene’s upstream conserved sequence

更新时间：2024-01-17

- Construction and identification of plasmid vector of microRNA on pneumocystis carinii major surface glycoprotein gene’s upstream conserved sequence
- Journal of Chongqing Medical University Vol. 35, Issue 2, Pages: 169-171(2010)
- 作者机构：
  
  重庆医科大学附属第一医院感染科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.02.007
  CLC： R346
- Published：2010，
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[1]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(02):169-171.

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 35(2):169-171(2010)
[1]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(02):169-171. DOI： 10.13406/j.cnki.cyxb.2010.02.007.

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 35(2):169-171(2010) DOI： 10.13406/j.cnki.cyxb.2010.02.007.

摘要

目的:构建和鉴定卡氏肺孢子菌（Pneumocystis carinii

PC）主要表面糖蛋白（Major surface glycoprotein

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

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。

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 expression was detected by RT-PCR and the fine structure of PC was observed by electronmicroscope.Results:Gel electrophoresis and DNA sequencing showed that the recombinant plasmid containing the correct and full microRNA oligonucleotide

and it could knock down the expression of mRNA in PC in vitro

it could also destroy PC.Conclusion:The microRNA plasmid

pPC-UCSm

was constructed successfully

and it could knock down the expression of PC in vitro.

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

Keywords

Pneumocystis cariniiMSG-UCSmicroRNAExpression plasmid

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Related Author

Fang Xiangdong

Wu Xianfeng

Ke Ben

姚云清

陈雅堂

马良

李文桂

唐滢浍

Related Institution

Department of Nephrology,the Second Affiliated Hospital of Nanchang University

Department of Infection Diseases,the First Affiliated Hospital,Chongqing Medical University

重庆医科大学附属第一医院感染科

Department of Orthopaedics,the Children’s Hospital,Chongqing Medical University

重庆医科大学附属儿童医院泌尿外科

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