Objective:To construct EGFP and HGF co-expression vector and to detect its expression in cultured eukaryocyte. Methods:A pair of primers of HGF with Sac I and BamH I were designed and constructed

then PCR was performed.The PCR products were inserted into cloning vector pMDlg-T and transferred into JM109.The PCR of colony

restriction analysis

and DNA sequencing were performed to identify the correctness of the recombinant.The HGF gene fragment obtained from pMD18-T/HGF that was digested with Sac I and BamH I

and then inserted into pIRES2-EGFP that was cut with Sac I and BamH I.The recombinant pIRES2-EGFP/HGF was identified with restriction analysis.The expression plasmid pIRES2-EGFP/HGF was transfected into HUVEC mediated by liposome reagent

then the expression of EFGP in cell were ob- served by fluorescence microscopy and HGF protein was detected with Western blotting.Result:The sequence of the cloned DNA fragment was identical to HGF that was reported on Gene bank

and the HGF gene was inserted into eukaryotic expres- sion vector pIRES2-EGFP correctly.The recombinant expression plasmid was successfully transferred into HUVEC observed by fluorescent microscope and effective expression of HGF was also testified by Western blotting.Conclusion:The recombinant eukaryotic co-expression vector of EGFP and HGF was successfully constructed and effectively expressed in HUVEC.