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

TAN Yan

doi:10.13406/j.cnki.cyxb.2008.03.005

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Construction and identification of plasmid vector of short interfering RNA 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 short interfering RNA on pneumocystis carinii major surface glycoprotein gene’s upstream conserved sequence
- Journal of Chongqing Medical University Issue 3, Pages: 264-266(2008)
- 作者机构：
  
  1. 重庆医科大学附属第一医院感染科
  2. 美国路易斯安那州新奥尔良大学医学院
  3. 重庆医科大学附属第一医院感染科,重庆,400016
  4. 路易斯安那州,70112
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2008.03.005
  CLC： R346
- Published：2008，
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[1]谭燕,姚云清,王莉妮,唐滢浍,李文桂,马良,陈雅棠.卡氏肺孢子主要表面糖蛋白UCS基因siRNA表达载体的构建和鉴定[J].重庆医科大学学报,2008(03):264-266.

TAN Yan. Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface glycoprotein gene’s upstream conserved sequence. [J]. Journal of Chongqing Medical University (3):264-266(2008)
[1]谭燕,姚云清,王莉妮,唐滢浍,李文桂,马良,陈雅棠.卡氏肺孢子主要表面糖蛋白UCS基因siRNA表达载体的构建和鉴定[J].重庆医科大学学报,2008(03):264-266. DOI： 10.13406/j.cnki.cyxb.2008.03.005.

TAN Yan. Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface glycoprotein gene’s upstream conserved sequence. [J]. Journal of Chongqing Medical University (3):264-266(2008) DOI： 10.13406/j.cnki.cyxb.2008.03.005.

摘要

目的:构建并鉴定艾滋病主要机会性感染病原体-卡氏肺孢子（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-UCS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

采用双酶切与测序法进行鉴定。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:成功构建和鉴定PC MSG-UCS基因的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’s upstream conserved sequence(UCS).Methods:Short hairpin RNA(shRNA) oligonucleotide targeting PC MSG UCS gene which was chemically synthesized and inserted into RNAi-Ready plasmid vector pTZU6+1 after annealing.The recombinant plasmid

PC-UCS

transformed into E.coil.TOP10 and amplified

was digested by restriction endonucleases HindⅢand 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-UCS

was constructed successfully.

关键词

PC MSG-UCSsiRNA表达载体

Keywords

Pneumocystis carinii MSG-UCSsiRNAexpression vector

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