目的：克隆人黑色素瘤相关抗原MAGE-C2的cDNA，构建真核、原核表达载体并转入相应细胞获得表达。方法:采用RT-PCR技术，从直肠癌细胞系SW480的总RNA中克隆了MAGE-C2 cDNA序列，并将开放读框分别插入质粒pEGFP-C1和pGEX-4T-3中，获得pEGFP-MAGE-C2绿色荧光蛋白（GFP）融合表达载体和pGEX-MAGE-C2谷胱甘肽转移酶（GST）融合蛋白表达载体，将pEGFP-MAGE-C2转染293T细胞，观察绿色荧光融合蛋白在细胞中的定位；将pGEX-MAGE-C2转化大肠杆菌BL21株，经IPTG诱导表达GST融合蛋白，并用谷胱甘肽亲和层析法纯化重组蛋白。结果:获得MAGE-C2开放读框并构建表达载体，测序结果与GenBank收录的序列相一致。真核表达载体转染293T细胞后显示MAGE-C2蛋白定位于细胞核；原核重组蛋白大部分以可溶性形式表达，亲和层析后，获得了较高纯度的MAGE-C2-GST融合蛋白。分析显示原核表达GST融合蛋白的相对分子质量为70 000，使用抗GST单克隆抗体进行Western blot分析证实为目的蛋白。结论:成功的获得MAGE-C2基因，构建了其表达载体并获得表达，为进一步制备MAGE-C2特异性单抗及深入探讨MAGE-C2可能参与肿瘤发生发展的机制提供了重要条件

Objective: To clone and express human MAGE-C2 gene and to investigate the expression pattern in transfected eukaryote cells. Methods: MAGE-C2 cDNA was amplified by RT-PCR from total RNA of human colorectal adnocarcinoma cell line SW480. Expression vectors of complete open reading frame (ORF) sequence of MAGE-C2 were constructed by PCR and gene cloning technique. After sequencing, the vectors were transfected into E. coli BL21 and 293T cell, respectively. Recombinant GST-MAGE-C2 fusion protein was expressed via IPTG induction. GST-MAGE-C2 fusion protein was purified through glutathione agarose column. Results:The sequence of cloned MAGE-C2 was identical with that reported in GenBank. GFP-MAGE-C2 fusion protein was localized on nuclear identified by fluorescent microscope. SDS-PAGE and Western blot analysis that purified GST-MAGE-C2 fusion protein exhibited a band with Mr. 70 000. Conclusion: The MAGE-C2 gene was cloned and expressed successfully, which not only provides the immunogen for further preparation of anti-MAGE-C2 antibodies, but also applies to research the mechanism of tumor's pathogenesis and cellular immunity response to MAGE.