Establishment of high efficient tranfection and expression system for phPPARy 1-1RES2-EGFP recombinant plasmid in vitro
- VernacularTitle:重组质粒phPPAR?1-IRES2-EGFP高效体外转染表达体系的建立
- Author:
Tao ZHANG
- Publication Type:Journal Article
- Keywords:
Peroxisome proliferator-activated receptory;
Transfection efficiency;
Eukaryotic expression;
293cells
- From:
Journal of Chongqing Medical University
2003;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish high efficient transfection and expression system in vitro for phPPAR?1-IRES2-EGFP re- combinant plasmid carrying human peroxisome proliferator-activated receptor?1(hPPAR?1)gene.Methods:Cationic lipid trans- fection reagent lipofectamine 2000 was used to transfect phPPAR?1-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 PPAR?1 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- PAR?1-IRES2-EGFP was(83?11)%.In transfected 293 cells,high efficient expression of hPPAR?1 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 phPPAR?1-IRES2-EGFP recombinant plasmid in vitro is successfully established,which provide a useful tool in investigating hPPAR?gene function as well as establishing molecular platform by which the candidates of unknown hP- PAR?ligands or activators can be found.
