Abstract:Objective:To construct the recombinant pEGFP-N1/hIL-1ra and test its transient expression in human gingival fibroblast cells, this may be useful of the treatment of periodontitis. Methods:Total RNA was extracted from human mammary cancer tissue. RT-PCR was performed to amplify the hIL-1ra encoding gene. Then PCR product was purified and cloned into pMD18-T. After DNA sequencing,both of the recombinant vector pMD18-T/hIL-1ra and plasmid pEGFP-N1 were digested with HindⅢ and BamHⅠ respectively. IL-1ra gene fragment was ligated into plasmid pEGFP-N1. The recombinant plasmid DNA was transformed into E.coli competent cells TOP 10 and positive clones were selected and tested. After being transfected by pEGFP-N1/hIL-1ra,the transient expression of IL-1ra gene in human gingival fibroblast(HGF) cells were detected by fluorescence microscope and RT-PCR. Results:hIL-1ra encoding gene fragment was 531 bp and was inserted into the eukaryotic expression vector pEGFP-N1/hIL-1ra correctly. And HGF cells,which were identified by both fluorescence microscope and RT-PCR,had a transient expression of IL-1ra after transfection. Conclusion:The new recombinant expression vector pEGFP-N1/hIL-1ra was constructed successfully and the HGF cells which could express IL-1ra transiently were also successfully made.