MAGE-1与人HSP70融合基因真核表达载体的构建及在小鼠黑色素瘤细胞内的表达  

Construction of MAGE-1 and human HSP70 fusion gene eukaryotic expression vector and its expression in mouse melanoma cells in vitro

作　　者：陈广生[1] 隋延仿[1] 叶菁[1] 宋宏萍[1] 黄亚渝[1] 马加海[1] 张秀敏[1] 

机构地区：[1]第四军医大学附属西京医院病理科,陕西西安710032

出　　处：《现代肿瘤医学》2006年第6期644-646,共3页Journal of Modern Oncology

基　　金：国家自然科学基金(30271464);全军医药卫生科研基金(01Z084)重点资助项目

摘　　要：目的:构建MAGE-1(melanomaantigen1)与人HSP70(heatshockprotein70)融合基因的真核表达载体PIRES2-EGFP-MAGE-1-HSP70,并观察在小鼠黑色素瘤细胞内的表达,为肿瘤DNA疫苗研究奠定基础。方法:HSP70基因经酶切后连入真核表达载体PIRES2-EGFP,再通过PCR方法扩增MAGE-1基因,测序后插入PIRES2-EGFP-HSP70中HSP70基因的5'端,构建了MAGE-1与人HSP70融合基因表达载体PIRES2-EGFP-MAGE-1-HSP70。结果:扩增了MAGE-1基因,测序结果表明与GenBank公布的序列一致,成功地构建了PIRES2-EGFP-MAGE-1-HSP70真核表达载体。结论:成功地构建MAGE-1与人HSP70融合基因的真核表达载体PIRES2-EGFP-MAGE-1-HSP70,并在小鼠黑色素瘤细胞系稳定表达,为进一步DNA肿瘤疫苗研究提供了实验基础。Objective：To construct the MAGE - 1 and human HSP70 fusion gene eukaryotic expression vector and the expression in mouse melanoma cells in vitro. Methods： MAGE - 1 genes were amplified by PCR. After sequen cing, the two genes were cloned into the eukaryotic expression vector PIRES2 - EGFP to construct fusion gene expres sion plnsmid PIRES2 - EGFP - MAGE - 1 - HSP70. Results： The MAGE - 1 gene were amplified successfully from plasmids, and their sequences were identical with that reported in GenBank. The fusion gene expression plasmid PIRES2 - EGFP - MAGE - 1 - HSP70 was successfully constructed. Conclusion： The MAGE - 1 and Hsp70 fusion gene eukaryotic expression vector PIRES2 - EGFP - MAGE - 1 - HSP70 was constructed which prepared the materials for the research of DNA vaccine .

关 键 词：黑色素瘤抗原1 人热休克蛋白70 融合基因 动物实验 

分 类 号：R730[医药卫生—肿瘤]