Establishment of high efficient tranfection and expression system for phPPARy 1-1RES2-EGFP recombinant plasmid in vitro

doi:10.13406/j.cnki.cyxb.2007.05.005

您当前的位置：

首页 >

文章列表页 >

Establishment of high efficient tranfection and expression system for phPPARy 1-1RES2-EGFP recombinant plasmid in vitro

更新时间：2024-01-17

- Establishment of high efficient tranfection and expression system for phPPARy 1-1RES2-EGFP recombinant plasmid in vitro
- Journal of Chongqing Medical University Issue 5, Pages: 459-463(2007)
- 作者机构：
  
  1. 重庆医科大学基础医学院药理学教研室
  2. 第三军医大学野战外科研究所分子生物学巾心
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2007.05.005
  CLC： R346
- Published：2007，
扫描看全文
[1]章涛,万敬员,杨俊卿,刘颖菊,周元国,周岐新.重组质粒phPPARγ1-IRES2-EGFP高效体外转染表达体系的建立[J].重庆医科大学学报,2007(05):459-463.

Establishment of high efficient tranfection and expression system for phPPARy 1-1RES2-EGFP recombinant plasmid in vitro. [J]. Journal of Chongqing Medical University (5):459-463(2007)
[1]章涛,万敬员,杨俊卿,刘颖菊,周元国,周岐新.重组质粒phPPARγ1-IRES2-EGFP高效体外转染表达体系的建立[J].重庆医科大学学报,2007(05):459-463. DOI： 10.13406/j.cnki.cyxb.2007.05.005.

Establishment of high efficient tranfection and expression system for phPPARy 1-1RES2-EGFP recombinant plasmid in vitro. [J]. Journal of Chongqing Medical University (5):459-463(2007) DOI： 10.13406/j.cnki.cyxb.2007.05.005.

摘要

目的:建立含人PPARγ1受体基因phPPARγ1-IRES2-EGFP重组质粒高效体外转染表达体系。方法:采用阳离子脂质体Lipofectamine 2000将phPPARγ1-IRES2-EGFP转染入293细胞

荧光显微镜观察转染细胞绿色荧光蛋白（GFP）报告基因表达强度及其转染效率

并对转染细胞hPPARγ1表达进行荧光定量PCR和Western Blot分析。结果:荧光显微镜下可见转染phPPARγ1-IRES2-EGFP质粒的293细胞GFP报告基因高表达

转染效率为（83±11）%;荧光定量PCR和Western Blot检测结果表明

转染细胞hPPARγ1表达水平高于空载体对照组3个数量级

说明导入的重组质粒能够高效表达hPPARγ1。结论:成功建立phPPARγ1-IRES2-EGFP重组质粒高效体外转染表达体系

为hPPARγ1受体功能研究和基于hPPARγ1为靶标的药物筛选平台建立打下了良好的基础。

Abstract

Objective:To establish high efficient transfection and expression system in vitro for phPPARγ1-IRES2-EGFP re- combinant plasmid carrying human peroxisome proliferator-activated receptorγ1(hPPARγ1)gene.Methods:Cationic lipid trans- fection reagent lipofectamine 2000 was used to transfect phPPARγ1-IRES2-EGFP recombinant plasmid and control pIRES2- EGFP plasmid to 293 cells.The profiles of GFP expression and transfection efficiency were measured by fluorescence mi- croscopy.Expression of PPARγ1 in transfected 293 cells were determined by werstern blot and real time quantitative poly- merase chain reaction.Results:GFP expression level was high in transfeced 293 cells

the transfection efficiency of phP- PARγ1-IRES2-EGFP was(83±11)%.In transfected 293 cells

high efficient expression of hPPARγ1 gene were detected both at mRNA and protein levels by real-time PCR and western blot analyses.Conclusion:High efficient transfection and expres- sion system for phPPARγ1-IRES2-EGFP recombinant plasmid in vitro is successfully established

which provide a useful tool in investigating hPPARγgene function as well as establishing molecular platform by which the candidates of unknown hP- PARγligands or activators can be found.

关键词

过氧化物酶体增殖物激活受体γl转染效率真核表达293细胞

Keywords

Peroxisome proliferator-activated receptorytransfection efficiencyEukaryotic expression293cells

references

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Eukaryotic expression of cis-inserted double copies of DuIFN-γ gene

Construction and expression of the pIRES-enchanced green fluorescent protein reporter gene vector carrying BMP₂ Gene

Construction of eukaryotic expression vector of open reading frame of unknown KH gene and its effect on proliferation of K562 cell

Construction of pcDNA3.1(+)/hFasL and its expression in COS-7 cells

Construction and verification of eukaryotic expression vector for mouse Hsp84

Related Author

Ieva Kotlarski

Allison Jilbert

et al

HUANG Ai-long

ZOU Haibo

et al

蒋电明

刘建伟

Related Institution

南澳州

澳大利亚阿得雷德大学微生物与免疫学系

Institute for Viral Hepatitis，Chongqing Medical University

重庆医科大学检验系

⁰