Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast / 重庆医科大学学报

ABSTRACT

Objective:To construct a vector containing human vascular endothelial growth factor,VEGF 121 (VEGF 121 ) with green fluorescence protein as the reporter gene,and then to observe its expression in osteoblast.Methods:VEGF 121 gene was amplified from foetus brain library by PCR,then ligated with pEGFP-C1 to form recombinant plasmid pEGFP-VEGF and transfected into osteoblast by liposome.The green fluorescence protein expression was observed by fluorescence microscopy and the VEGF expression was last detected by immunocytochemistry and ELISA.Results:VEGF 121 gene fragment was amplified by PCR and ligated to pEGFP-C1,then transfected into osteoblast successfully.The strong expression of green fluorecensce protein was observed by fluorecensce microscopy,and the expression of VEGF protein was detected by immunocytochemistry and ELISA.Conclusion:Recombinant plasmid pEGFP-VEGF is effectivly expressed after being transfected into osteoblast in vitro,which provides theoretical support for further study with its simple and reliable detecting methods.