Construction and identification of eukaryotic expression vectors of soluble HLA-A2 and human IgG1-Fc fragment

Objective To construct a recombinant plasmid pcDNA3.0-sHLA-A2-IgG1-Fc,and to provide a firm basis on construction of soluble HLA-A2-IgG1-Fc protein by eukaryotic expression system. Methods Total cell RNA was extracted from the cell line T2 and the cDNA sequences with encoding signal peptide were amplified by RT-PCR.The cDNA fragment was inserted into the eukaryotic expression vector pcDNA3.0 and the recombinant plasmid was identified by restriction endonuclease digestion and sequencing.The gene of objective IgG1-Fc fragment was amplified from plasmid PIG by PCR.After restriction endonuclease digestion,the cDNA frogment was inserted into the constructed vector pcDNA3.0-sHLA-A2 and the recombinant plasmid was identified by resriction endonuclease digestion and sequencing.Results After restriction endonuclease treatment and sequencing,it was confirmed that HLA-A2 and IgG1-Fc has been inserted into vector pcDNA3.0. Conclusion The recombinant plasmid pcDNA3.0-sHLA-A2-IgG1-Fc has been constructed successfully.