[Establishment of mouse melanoma B16 cell lines co-expressing MAGE-1 and EGFP].
Guangsheng Chen, Jing Ye, Hongping Song, Xiumin Zhang, Ya-Yu Huang, Jiahai Ma, Yan-Fang Sui
Abstract
Guangsheng Chen, Jing Ye, Hongping Song, Xiumin Zhang, Ya-Yu Huang, Jiahai Ma, Yan-Fang Sui
Abstract
AIM: To construct the melanoma antigen-1(MAGE-1) eukaryotic expression plasmid and express MAGE-1 in mouse melanoma B16 cells. METHODS: The MAGE-1 gene was amplified by PCR and cloned into the eukaryotic expression vector pIRES2-EGFP to construct the pIRES2-EGFP-MAGE-1 plasmid. The plasmid was transfected into the B16 cells. The EGFP expression was detected under fluoroscent microscope and the MAGE-1 expression was detected by immunohistochemistry staining. RESULTS: The eukaryotic expression vector pIRES2-EGFP-MAGE-1 was constructed and transfected successfully into B16 cells, and the EGFP and MAGE-1 genes were co-expressed in the B16 cells. CONCLUSION: A mouse melanoma cell line B16 co-expressing MAGE-1 and EGFP genes has been established successfully, which lays the foundation for the research on application of MAGE-1 in the tumor immunotherapy.
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AIM: To construct the melanoma antigen-1(MAGE-1) eukaryotic expression plasmid and express MAGE-1 in mouse melanoma B16 cells. METHODS: The MAGE-1 gene was amplified by PCR and cloned into the eukaryotic expression vector pIRES2-EGFP to construct the pIRES2-EGFP-MAGE-1 plasmid. The plasmid was transfected into the B16 cells. The EGFP expression was detected under fluoroscent microscope and the MAGE-1 expression was detected by immunohistochemistry staining. RESULTS: The eukaryotic expression vector pIRES2-EGFP-MAGE-1 was constructed and transfected successfully into B16 cells, and the EGFP and MAGE-1 genes were co-expressed in the B16 cells. CONCLUSION: A mouse melanoma cell line B16 co-expressing MAGE-1 and EGFP genes has been established successfully, which lays the foundation for the research on application of MAGE-1 in the tumor immunotherapy.
Key concepts: Transfection, Green fluorescent protein, Molecular biology, Plasmid, Melanoma, Cell culture, Gene, Biology