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目的:从肝癌组织中克隆肿瘤相关抗原基因MAGE-1的全长cDNA,为该基因及其编码抗原应用于肝癌的免疫治疗奠定基础.方法:根据MAGE-1编码区上、下游序列设计一对引物,用RT-PCR方法从肝癌组织中扩增该基因的全长cDNA,酶切产生粘性末端后连接入PUC19质粒.经初步酶切鉴定后,通过DNA序列分析获取所克隆基因片断的核苷酸序列.结果:成功地克隆了MAGE-1基因的全长cDNA,同时还获得一约750bp的MAGE-3基因片段及一与MAGE-6和MAGE-12高度同源的基因的克隆.此与MAGE-6,-12高度同源的基因可能为一未知MAGE家族成员.结论:肝癌组织中表达多种MAGE家族基因,甚至可能存在未知MAGE家族基因的表达,这将为寻找肝癌免疫治疗的攻击靶点开辟新的途径.
Objective: To clone the full-length cDNA of tumor-associated antigen gene MAGE-1 from hepatocellular carcinoma and lay a foundation for the application of this gene and its encoded antigen in liver cancer immunotherapy. Methods: Design a pair of sequences based on the upstream and downstream sequences of the MAGE-1 coding region. The primers were used to amplify the full-length cDNA of this gene from liver cancer tissue by RT-PCR method, and the sticky end was cut into the PUC19 plasmid. After preliminary enzyme digestion, the nucleoside of the cloned gene fragment was obtained by DNA sequence analysis. Acid sequence. Results: The full-length cDNA of the MAGE-1 gene was successfully cloned and a 750 bp MAGE-3 gene fragment and a clone of a gene highly homologous to MAGE-6 and MAGE-12 were also obtained. MAGE-6,-12 highly homologous gene may be an unknown MAGE family member. Conclusion: Many MAGE family genes are expressed in hepatocellular carcinoma tissues, and even the expression of unknown MAGE family genes may exist. This will be an attempt to find an immunotherapy for liver cancer. Targets open up new avenues.