Eukaryotic expression and identification of human DC-SIGN and EGFP fusion protein / 中国免疫学杂志

ABSTRACT

Objective:To construct the eukaryotic vector of human DC-SIGN and EGFP fusion protein,and to identify the protein in cell line COS7.Methods:RT-PCR,T-vector and pEGFP-C1 vector were used to construct the recombinant expressing plasmid encoding for the fusion protein of DC-SIGN and EGFP.COS7 cells were transfected with the plasmid.Real-time PCR and laser scanning confocal microscope were used to quantificate expression of the fusion protein and cell function of uptaking BCG.Results:DC-SIGN cDNA was successfully cloned into the eukaryotic vector pEGFP-C1.The recombinant vector was transfected into COS7,real-time PCR test showed the amount of mRNA encoding for the fusion protein was 4.52?1011 copies/ml and laser scanning confocal microscope confirmed that the cells could uptake BCG.Conclusion:We have constructed a recombinant vector expressing DC-SIGN and EGFP fusion protein.