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

WANG Li-ni

doi:10.13406/j.cnki.cyxb.2008.05.005

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

更新时间：2024-01-17

- Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface gene
- Journal of Chongqing Medical University Issue 5, Pages: 517-519(2008)
- 作者机构：
  
  1. 重庆医科大学附属第一医院感染科
  2. 重庆医科大学附属第一医院感染科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2008.05.005
  CLC： R346
- Published：2008，
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[1]王莉妮,姚云清,谭燕,唐滢浍,李文桂,马良,陈雅堂.卡氏肺孢子表面糖蛋白siRNA表达载体的构建和鉴定[J].重庆医科大学学报,2008(05):517-519+525.

WANG Li-ni. Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface gene. [J]. Journal of Chongqing Medical University (5):517-519(2008)
[1]王莉妮,姚云清,谭燕,唐滢浍,李文桂,马良,陈雅堂.卡氏肺孢子表面糖蛋白siRNA表达载体的构建和鉴定[J].重庆医科大学学报,2008(05):517-519+525. DOI： 10.13406/j.cnki.cyxb.2008.05.005.

WANG Li-ni. Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface gene. [J]. Journal of Chongqing Medical University (5):517-519(2008) DOI： 10.13406/j.cnki.cyxb.2008.05.005.

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。

Abstract

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 HindⅢand EcoRⅠand identified by gelelectrophoresis 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-MSG was constructed successfully.

关键词

卡氏肺孢子主要表面球蛋白siRNA表达载体

Keywords

Pneumocystis cariniiMajor surface glycoproteinsiRNAExpression plasmid

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