Expression of Truncated uPA Fused to EGFP in HEK293F Cells

Objective: To construct and express eukaryotic expression vectors of the truncated urokinase-type plas minogen activator(uPA) fused to enhanced green fluorescent protein(EGFP).Methods: EGFP and truncated uPA genes were amplified by PCR using plasmid pIRES2-EGFP and recombinant plasmid pcDNA3.1(+)/uPA as tem plates,and inserted into eukaryotic expression vector pcDNA3.1(+) sequentially.The constructed recombinant plas mids were transfected into HEK293F cells,and treated with high concentration G418.The expression of recombi nant proteins was detected by confocal microscopy and ELISA.Results: DNA sequencing proved that the eukaryot ic expression vectors of the fusion proteins were constructed successfully.And the green fluorescent protein could be observed in cells by confocal microscopy after the transfection,and the stable expression cell lines were got af ter selected by G418.ELISA showed that the secreting type fusion proteins exist in supernatant.Conclusion: Re combinant plasmids have been constructed and expressed in HEK293T cells,which will contribute to further re search of the interaction of uPA and its biological function in cells.