[Establishment of mouse melanoma B16 cell lines co-expressing MAGE-1 and EGFP].
OBJECTIVE
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.


CONCLUSIONS
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.
Chen
Guang-sheng
GS
Department of Pathology, Fourth Military Medical University, Xi'an 710032, China. chengs2001@sina.com
Ye
Jing
J
Song
Hong-ping
HP
Zhang
Xiu-min
XM
Huang
Ya-yu
YY
Ma
Jia-hai
JH
Sui
Yan-fang
YF
D000818:Animals; D000951:Antigens, Neoplasm; D045744:Cell Line, Tumor; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008546:Melanoma, Experimental / Q000378:metabolism* / Q000473:pathology; D058950:Melanoma-Specific Antigens; D051379:Mice; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids; D014162:Transfection
D000951:Antigens, Neoplasm; C072011:MAGEA1 protein, human; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins