Objective    To clone Ang Ⅱ type 2 receptor (AT2R) gene

obtain enhanced green fluorescent protein(EGFP) vector for the gene

and then observe its expression in primary vascular smooth muscle cells (VSMCs) after transfection.     Methods     Utilizing plasmid pUHD-10.3/AT2R as a template

the full length cDNA fragment of AT2R was amplified by polymerase chain reaction (PCR). Eukaryotic expression vector pEGFP-N2/AT2R was constructed by cloning AT2R gene into vector pEGFP-N2. After transfecting AT2R into primarily cultured VSMCs

invert fluorescent microscopy was employed to observe VSMCs growth as well as AT2R expression in VSMCs. Transfection efficiency of AT2R in VSMCs was evaluated by image analyzor

the protein and mRNA expressions of AT2R in VSMCs were examined by Western blotting and RT-PCR.     Results     Eukaryotic expression vector pEGFP-N2/AT2R was successfully constructed. The eukaryotic vector expressing AT2R gene was efficiently transfected and expressed in VSMCs. The transfection efficiency of AT2R gene in VSMCs was about 40%. RT-PCR and Western blotting indicated that AT2R was expressed in cultured VSMCs at mRNA and protein levels.     Conclusion       cDNA sequence encoding AT2R gene is successfully cloned into pEGFP-N2 which can be efficiently expressed in primary VSMCs.