Objective:To construct and identify pEGFP-HDGF over expression plasmid in U87 cells. Methods:HDGF fragment was amplified by RT-PCR method and then inserted into pEGFP-N1 vector. The level of HDGF was identified by RT-PCR and western blot after transfection of positive recombinant plasmid into glioma cell line U87. The biological activity of HDGF-GFP fusion protein was tested by the proliferation assay. Results:Fluorescence microscopy demonstrated that pEGFP-HDGF was successfully transfected into U87 cells. Over expression of HDGF was confirmed by RT-PCR and western blot. The proliferation rate of U87 cell transfected with pEGFP-HDGF plasmid was significantly higher than that of negative control. Conclusion:pEGFP-HDGF over expression plasmid with biological activity was successfully constructed and stablely transfected into U87 cells.