中文摘要：

目的：构建真核表达载体，并在小鼠黑色素瘤B16细胞中稳定表达。方法：用PCR方法扩增质粒pUC19-MAGE-1中目的基因MAGE-1，连接到真核表达载体pCDNA3．1中，构建真核表达pCDNA3．1-MAGE-1，脂质体法转染小鼠黑色素瘤B16细胞。G418筛选阳性克隆（B16-MAGE-1），用RT—PCR和Western blot检测阳性克隆中mRNA和蛋白质的表达；分别以2×10^5个B16细胞和B16-MAGE-1接种于C57BL／6小鼠右侧背部皮下，观察B16-MAGE-1细胞的致瘤能力。结果：用PT—PCR与Western bolt分别在B16-MAGE-1细胞中检测到人MAGE-1基因的mNRA和蛋白质的表达，两组共20只小鼠均皮下成瘤，且肿瘤大小没有明显差异。结论：成功构建了真核表达载体pCDNA3．1-MAGE-1，获得了稳定表达人MAGE-1基因的小鼠黑色素瘤B16细胞系，并保持很好的致瘤能力，为研究MAGE家族在肿瘤免疫治疗中的应用奠定了基础。

英文摘要：

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 fight 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.