Construction of recombinant pEGFP-heNOS plasmid and its expression in EPC

Objective To construct a recombinant plasmid containing an enhanced green fluores- cent protein (EGFP) reporter gene for the role of human endothelial nitric oxide synthase (heNOS),and observe its expression in endothelial progenitor cells (EPCs) derived from human bone marrow.Methods Recombinant plasmid pEGFP-heNOS was constructed by techniques of gene recombination,and trans- feeted into EPCs by liposome.The GFP expression was observed by fluorescence microscopy and the heNOS expression in EPCs was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot.Results Recombinant plasmid pEGFP-heNOS was constructed and transfected into EPCs successfully.The strong expression of GFP was observed by fluorecensce microscopy.The heNOS mRNA and protein could be detected within the transfected ceils by RT-PCR and Western blot respec- tively.Conclusion Recombinant plasmid pEGFP-heNOS is effectively expressed after being transfected into EPCs in vitro,which provides theoretical support for further gene therapy.