Establishment of mouse melanoma B16 cell lines which stably express human MAGE-1

Objective:To construct the eukaryotic expression vector which can be expressed in B16 cells. Methods: The MAGE-1 gene was amplified by PCR from pUC19-MAGE-1,then cloned into pCDNA3.1 to construct the pCDNA3.1-MAGE-1 plasmid. Under the mediation of lipofectamine, the plasmid was transfected into the mouse melanoma B16 cells, and then the positive clones were selected by G418.The expression of MAGE-1 mRNA and protein were respectively detected by RT-PCR and Western Blot. Mice (10 per group) were challenged with the B16 cells and B16-MAGE-1 cells (2×10~5 cells/mouse, respectively) in the right back. Results: The plasmid was constructed and transfected into B16 cells successfully. The expression of mRNA and protein of MAGE-1 were approved in the positive clones,two groups both grow melanoma. Conclusion: The eukaryotic expression plasmid pCDNA3.1-MAGE-1 was constructed successfully. The B16 cell line that stably expressed MAGE-1 is obtained, which will contribute to the research of MAGE family in the immunotherapy of tumor.