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 Sal Ⅰ and BamH Ⅰ , 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. Key words: Dendritic cell-specific ICAM-3 grabbing nonintegrin; 293T cells; Protein expression