Vector construction and eukaryotic expression of the human DC-SIGN

Objective To constructed recombinant eukaryotic plasmid VRC4409-DC-SIGN,then to express the protein DC-SIGN in 293T cells.Methods Dendritic cells(DC) was obtained from the human blood culture.DC-SIGN gene was amplified from cDNA of DC.The DC-SIGN gene and vector VRC4409 were digested by SalⅠand BamHⅠ,and the DC-SIGN gene fragments and the plasmids of VRC4409 were ligated with T4 DNA rapid ligase to form the recombinants VRC4409-DC-SIGN.After restriction analysis and sequencing,the plasmid VRC4409-DC-SIGN was transfected into 293T cells in the mediation of liposome.The expression of DC-SIGN was analyzed by western blot.Results The recombinant plasmids VRC4409-DC-SIGN were confirmed by restriction enzyme assay and sequencing.The DC-SIGN protein were successfully expressed in 293T cells.Conclusions The eukaryotic expression vector VRC4409-DC-SIGN was correctly constructed and the DC-SIGN protein was successfully expressed in 293T cells.This will facilitate the following study on DC-SIGN.