Construction of RNA interference expression vectors of human Hsp90 alpha

NIE Bin; et al

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Construction of RNA interference expression vectors of human Hsp90 alpha

更新时间：2024-01-17

- Construction of RNA interference expression vectors of human Hsp90 alpha
- Journal of Chongqing Medical University Issue 6, Pages: 597-600(2007)
- 作者机构：
  
  1. 重庆医科大学检验系临床检验诊断学省部共建教育部重点实验室重庆市重点实验室
  2. 重庆医科大学检验系临床检验诊断学省部共建教育部重点实验室重庆市重点实验室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC： R346
- Published：2007，
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[1]聂滨,曾照芳.Hsp90α基因RNAi的载体构建研究[J].重庆医科大学学报,2007(06):597-600.

NIE Bin, et al. Construction of RNA interference expression vectors of human Hsp90 alpha. [J]. Journal of Chongqing Medical University (6):597-600(2007)
[1]聂滨,曾照芳.Hsp90α基因RNAi的载体构建研究[J].重庆医科大学学报,2007(06):597-600. DOI：

NIE Bin, et al. Construction of RNA interference expression vectors of human Hsp90 alpha. [J]. Journal of Chongqing Medical University (6):597-600(2007) DOI：

摘要

目的:构建Hsp90α（Heat shock protein 90 alpha）siRN（A Small interfencing RNA）表达载体

为后续的功能研究提供平台。方法:根据Hsp90α基因cDNA序列

设计有小发夹结构的3条寡核苷酸序列到空载体Psuper EGFP1中

转化TOP10菌株

提取质粒

进行酶切鉴定和测序分析。将三条序列和对照序列的质粒分别转染HUK293细胞

提取总RNA

用RT-PCR检测Hsp90αmRNA的量。结果:经酶切鉴定、电泳证明

含作用序列的重组质粒pSuper-Hsp90α1

pSuper-Hsp90α2

及pSu-per-Hsp90α3构建是成功的

重组质粒pSuper-Hsp90α1

pSuper-Hsp90α2测序分析完全正确。用RT-PCR初步证明三条均有干扰效果

pSuper-Hsp90α1最明显。结论:用于Hsp90αRNAi的重组质粒可于体外成功构建

初步确定pSuper-Hsp90α1为最佳作用靶点。

Abstract

Objective:To construct the expression vector pSuper.Hsp90α/pSuper for further research of the Hsp90α.Methods: In accordance with the Hsp90α sequence in gene bank

three sequences respectively targeting Hsp90α were cloned into the RNA expression vector pSuper.Results:It was confirmed by enzyme digestion

and electrophoresis that the construction of the recombinant expression vector pSuper-Hsp90α1

pSuper-Hsp90α2 and pSuper-Hsp90α3 was successfu1.pSuper-Hsp90α1 and pSuper-Hsp90α2 were successfu1 by sequencing.The expression of Hsp90α mRNA in HEK293 cells with pSuper-Hsp90α1

pSuper-Hsp90α2 and pSuper-Hsp90α3 decreased

and pSuper-Hsp90α1 notably decreased.Conclusion:The recombinant plasmid against Hsp90α RNAi can be successfully constructed in vitro

and pSuper-Hsp90α1 is the possible optimal target.

关键词

RNA干扰热休克蛋白90α亚型

Keywords

RNA interferenceHsp90α

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Related Institution

Department of Radiology,The First Affiliated Hospital of Chongqing Medical University

Department of Emergency and Intensive Care Unit,The First Affiliated Hospital of Chongqing Medical University

Teaching and Research Section of Pathology,College of Basic Medicine,Molecular Medicine and Cancer Research Center,Chongqing Medical University

重庆医科大学公共卫生学院生殖医学教研室

Department of Biochemistry and Molecule Biology,College of Basic Medicine,Chongqing Medical University

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