Construction and verification of a specific GFP expression vector driven by promoter of human RIP2 gene

Aim:To construct a specific GFP expression vector driven by promoter of human RIP2 gene. Methods: Two different DNA segments of RIP2 gene promoter were amplified by PCR from human genome DNA and correctly connected to the vector pEGFP-C2 which had cut out promoter by restriction enzyme.Plasmids were identified by VspⅠand NheⅠ double digestion,and sequence analysis.Recombinant plasmids were transfected into cell line HEK293 by JetPeiTM.At 48 hours post-transfection,cells were observed under the inverted fluorescence microscope.Results: Two recombinant plasmids were the same as the design confirmed by restriction digestion and sequence analysis.The results of cell transient transfection indicated that the GFP could be observed in HEK293 by two plasmids transfection.The GFP expression of constructed pEGFP-C2-RIP2(750 bp)wt was stronger than that of pEGFP-C2-RIP2(941 bp)wt.Conclusion:The GFP expression vector driven by human RIP2 gene promoter is successfully constructed.