Construction And Identification Of PPARγ-Overexpression Cells

To study effect of PPARγ on atherosclerosis, by establishing a stable transfected cell overexpression model for PPARγ to investigate the interaction with its Specific ligands and the mechanism involved in formation. In this research, human PPARγgene fragment was obtained from human macrophage by nest RT-PCR and subcloned into eukaryotic overexpression plasmid. After sequencing, the recombinant plasmid was transfected into 293 T cells by lipidosome. The expression and activity of exogenous PPARγin 293 T cells was detected by western-blot and Luciferase reporter. After all the experments above, the cloned ORF of PPARγ gene fragment was confirmed by DNA sequencing and the subcloned recombinant overexpression plasmid was in right position. After 48 h transfection, abundant PPARγ was expressed. The results indicated that high expression PPARγ cell was established which could be used for further studies and provides powerful tool for PPARγfunction studying.