TRAIL真核表达质粒的构建及其在膀胱癌细胞中的表达  

Construction and identification of eukaryotic expression vector of human TRAIL gene

作　　者：林海群[1] 王绍勇[2] 李宝生[1] 翟利民[1] 

机构地区：[1]山东省肿瘤医院,山东济南250117 [2]山东大学第二医院

出　　处：《山东医药》2008年第45期13-15,共3页Shandong Medical Journal

摘　　要：目的构建TRAIL真核表达质粒并初步研究其在膀胱癌细胞中的表达。方法提取Jurkat淋巴瘤细胞总RNA,进行RT-PCR反应,产物测序证实后插入经同样酶切的质粒PcDNA3.1中,构建成真核表达质粒PcD-NA3.1-TRAIL。Western blot法鉴定其能否在膀胱癌细胞中表达。结果Jurkat淋巴瘤细胞的RT-PCR产物经测序证实为TRAIL基因的全长序列;经酶切、测序证实成功构建PcDNA3.1-TRIAL质粒;Western blot结果显示,转染上述质粒的膀胱癌细胞中有TRAIL的表达。结论成功构建了表达全长TRAIL的真核表达载体,并通过Westernblot验证其可在膀胱癌细胞中表达。Objective To construct TRAIL gene eukaryotic expression vector pcDNA3.1-TRAIL and stuty its exspression in human bladder cancer lion. Methods TRAIL gene was amplified from the mRNA of Jurkat cells by RT-PCR. PCR product was digested, and cloned intoeukaryotic expression vector pcDNA3.1 was digested by the same enzymes. Plasmid pcDNA3.1-TRAILwas transferred into human bladder cancer cells and Western blot detected its expression. Results cDNA was amplified by RT-PCR and the sequencing result proved that it was TRAIL full length. Western Blot could detect expression of TRAIL in bladder cancer cells. Conclusion The eukaryotic expression vector pcDNA3.1 TRAIL has been constructed successfully and expresses the correct protein in bladder cancer cells.

关 键 词：TRAIL 质粒 膀胱肿瘤 

分 类 号：R737.14[医药卫生—肿瘤]