Construction and expression of recombinant gene vecter with fusion of soluble CD40 and EGFP in dendritic cells

Objective To construct a recombinant plasmid carrying enhanced green fluorescent protein(EGFP) and mouse soluble CD40 gene(sCD 40) and detect its expression levels in dendritic Cells (DC 2.4).Methods The standard cloning technology was employed to construct the EGFP/ VEGF fusion gene which was identified with double enzyme digestion and DNA sequencing,then this recombinant plasmid was transfected into mouse dendritic Cells (DC2.4) with lipofectamine.The expression levels of the fusion gene were detected by fluorescence microscope,fluorescence spectrophotometer,SDS-PAGE,and flow cytomerey (FCM),respectively.Results The fusion gene was observed in transiently transfected dendritic cells.The fusion protein was secreted into supernatant.These data suggested that the EGFP tag did not interfere with the natural assembly and the biological activity of soluble CD40.Conclusions The recombinant plasmid carrying enhanced green fluorescent protein and mouse soluble CD40 gene is successfully constructed and expressed positively in rat DCs with activity. Key words: Soluble CD40 ;  Green fluorescent protein ;  Fusion proteins ;  Gene expression ;  Dendritic cells