Construction and expression of recombinant human eukaryotic expression vector pMAGE-A1/EGFP

Objective To clone MAGE-A1 gene open reading frame and construct eukaryotic expression vector with green fluorescent protein, and to transfect human PBMC-derived DCs. Methods MAGE-A1 open reading frame was amplified from Mel-526 by RT-nest PCR with specific primers, and then cloned into eukaryotic expression vector with EGFP. After construction of recombinant plasmid pMAGE-A1/ EGFP, human PBMC-derived DCs were transfected with pMAGE-A1/EGFP by electroporation, and protein expression was identified by fluorescent microscopy. Results The pMAGE-A1/EGFP eukaryotic expression vector was constructed successfully. MAGE-A1 and EGFP were expressed after transfection into DCs. Conclusion The pMAGE-A1/EGFP eukaryotic expression vector will provide a new modality of tumor immunotherapy targeting MAGE-A1 gene.