中文摘要：

目的：建立phPPARδ-IRES2-EGFP重组质粒高效体外转染表达的体系。方法：采用阳离子脂质体Lipofectamine2000将phPPARδ-IRES2-EGFP转染入293细胞,荧光显微镜观察质粒转染细胞GFP报告基因表达强度及转染效率,并对转染细胞hPPARδ的表达进行荧光定量PCR和Western Blot检测。结果：荧光显微镜下可见转染phPPARδ-IRES2-EGFP质粒的293细胞绿色荧光蛋白（GFP）报告基因高表达,转染效率高达（85±10）%;荧光定量PCR和Western Blot检测的结果表明,phPPARδ-IRES2-EGFP转染的293细胞其hPPARδ表达水平高于空载体转染对照组（P〈0.01）。结论：成功建立phPPARδ-IRES2-EGFP质粒的高效体外转染表达体系,为hPPARδ受体功能的研究及基于hPPARδ为靶标的药物筛选平台的建立奠定了基础。

英文摘要：

Objective： To establish high efficient transfection and expression system in vitro for phPPARδ- IRES2-EGFP recombinant plasmid carrying human peroxisome proliferator-activated receptor δ（phPPARδ） gene. Methods： Cationic lipid transfection reagent lipofectamine 2000 was used to transfect phPPARδ- IRES2-EGFP recombinant plasmid and control pIRES2-EGFP plasmid to 29δ cells. The profiles of GFP expression and transfection efficiency were measured by fluorescence microscopy. Expression of PPARδ in transfected 29δ cells was determined by werstern blot and real time quantitative polymerase chain reaction （PCR）. Results. GFP expression level was high in transfeced 29δ cells, the transfection efficiency of phPPARδ-IRES2-EGFP was （85±10）%. High efficient expression of phPPARδ gene was detected in transfected 29δ ceils both at mRNA and protein levels by real-time PCR and western blot analyses. Conclusion： Successful establishment of high efficient tranfection and expression system for phPPARδ-IRES2-EGFP recombinant plasmid in vitro which provide a useful tool in investigating phPPARδ gene function as well as establishing molecular platform by which the unknown phPPARδ ligands can be found.