| 人MAGE-3真核表达载体的构建与表达 |
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| 引用本文： | 吴金民,孙晓东,刘杏娥.人MAGE-3真核表达载体的构建与表达[J].实用肿瘤杂志,2003,18(4):271-274. |
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| 作者姓名： | 吴金民 孙晓东 刘杏娥 |
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| 作者单位： | 浙江大学医学院附属邵逸夫医院肿瘤中心,浙江大学邵逸夫临床医学研究所,浙江,杭州,310016 |
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| 摘 要： | 目的 克隆人黑色素瘤特异性抗原MAGE-3基因，构建真核表达载体，建立MAGE-3阳性细胞株，制备肿瘤核酸疫苗。方法 用RT—PCR方法扩增MAGE-3cDNA，以pcDNA3．1 为载体，构建重组表达质粒pcDNA3．1／MAGE-3。重组质粒用脂质体转染鼠B16细胞，经RT—PCR、细胞免疫染色及免疫印迹法鉴定转化细胞中MAGE-3的表达。结果 正确构建了pcDNA3．1／MAGE-3重组质粒，并且在转化细胞中检测到了MAGE-3的表达。结论 成功构建了pcDNA3．1／MAGE-3真核表达质粒，建立了稳定表达人MAGE-3的鼠B16细胞株。
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| 关 键 词： | MAGE-3 真核表达载体 构建 表达 黑色素瘤 |
| 文章编号： | 1001-1692(2003)04-0271-04 |
| 修稿时间： | 2002年11月11 |
Construction of human MAGE-3 eukaryotic expression plasmid and its expression in vitro |
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| WU Jin min,SUN Xiao dong,LIU Xing e.Construction of human MAGE-3 eukaryotic expression plasmid and its expression in vitro[J].Journal of Practical Oncology,2003,18(4):271-274. |
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| Authors: | WU Jin min SUN Xiao dong LIU Xing e |
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| Abstract: | Objective To clone human MAGE 3 gene, to construct a eukaryotic expression vector and to transfect it into a human MAGE 3 B16 cell lines. Methods The recombinant eukaryotic expression plasmid pcDNA3 1/MAGE 3 was constructed by ligating MAGE 3 gene, which was amplified by RT PCR, and the pcDNA3 1+ vector. The recombinant plasmids were transfected into B16 cells by liposome, the expression of MAGE 3 was verified by RT PCR, immunocytochemistry and Western blot. Results The recombinant pcDNA3 1/MAGE 3 plasmid was constructed successfully and the expression of MAGE 3 gene in transfected B16 cells was confirmed. Conclusion The recombinant mammalian expression plasmid has been successfully constructed and a B16 hMAGE 3 cell line established, which expresses MAGE 3 protein stably |
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