Construction of eukaryotic expression plasmid experssing shRNA targeting EGFR gene family

HUANG Huan; et al

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Construction of eukaryotic expression plasmid experssing shRNA targeting EGFR gene family

更新时间：2024-01-17

- Construction of eukaryotic expression plasmid experssing shRNA targeting EGFR gene family
- Journal of Chongqing Medical University Issue 4, Pages: 399-402(2008)
- 作者机构：
  
  1. 重庆医科大学附属第一医院肿瘤科
  2. 重庆市肿瘤研究所
  3. 重庆医科大学基础医学院放射医学教研室,重庆,400016
  4. ,重庆,400030
- 作者简介：
- 基金信息：
- DOI：
  CLC： Q78
- Published：2008，
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[1]黄环,吴永忠,李少林,郭启帅,林远洪,彭志平.RNA干扰EGFR基因家族真核表达载体的构建[J].重庆医科大学学报,2008(04):399-402.

HUANG Huan, et al. Construction of eukaryotic expression plasmid experssing shRNA targeting EGFR gene family. [J]. Journal of Chongqing Medical University (4):399-402(2008)
[1]黄环,吴永忠,李少林,郭启帅,林远洪,彭志平.RNA干扰EGFR基因家族真核表达载体的构建[J].重庆医科大学学报,2008(04):399-402. DOI：

HUANG Huan, et al. Construction of eukaryotic expression plasmid experssing shRNA targeting EGFR gene family. [J]. Journal of Chongqing Medical University (4):399-402(2008) DOI：

摘要

目的:构建针对表皮生长因子受体（Epidermal growth factor recepter

EGFR）家族的小分子发夹状RN（A Small hairpinRNA

shRNA）干扰质粒表达载体

为本课题下一步研究提供有力工具。方法:选择放疗不敏感的卵巢癌细胞SKOV3中高表达的EGFR家族同源性基因为靶基因

根据shRNA的设计原则

设计实验组及对照组shRNA。体外合成2段编码短发夹RNA序列的DNA单链

退火

克隆到相应载体pGensil-1的polⅢ启动子下游。结果:经酶切和DNA测序鉴定成功构建了EGFR家族同源性基因的RNA干扰表达载体

分别命名为pGensil-1-E及pGensil-1-C。为下一步体外、体内实验奠定了基础。

Abstract

Objective:To construct the eukaryotic expression plasmid expressing shRNA targeting EGFR gene family as a tool for following experiments in vitro and in vivo.Methods:EFGR family highly expressed in low radiosensitive ovarian cancer cells SKOV3

chosen highly expressed homologous genes in EFGR family as targeted gene.According to guidelines for shRNA design

designed experimental shRNA and negative control shRNA.In vitro synthesized 2 of the coding sequence of short hairpin RNA DNA single-strands

annealing

and cloned into the corresponding vector pGensil-1 of pol Ⅲ promoter downstream

named pGensil-1-E and pGensil-1-C.Confirmed by restrict endonuclease digestion and DNA sequencing.Conclusions:Eukaryotic expression plasmid experssing shRNA targeting homologous genes in EFGR family was constructed successfully.

关键词

EGFR家族RNA干扰真核细胞表达载体

Keywords

EFGR familyRNA interferenceEukarvotic expression vector

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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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