Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing, 210046 Jiangsu, People's Republic of China.
J Fish Biol. 2011 Sep;79(3):747-60. doi: 10.1111/j.1095-8649.2011.03058.x. Epub 2011 Aug 11.
In this study, the complementary DNA (cDNA) of Japanese pufferfish Takifugu rubripes tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) was cloned by reverse-transcription PCR. The open reading frame of the TRAIL consisted of 870 bases. The deduced amino-acid sequence of the TRAIL showed a high homology with the sequences of other teleosts. Recombinant soluble TRAIL was fused with a small ubiquitin-related modifier gene to enhance the soluble expression level in Escherichia coli BL21 (DE3). In vitro, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide (MTT) assay indicated that the purified soluble TRAIL was able to induce apoptosis of Jurkat and HeLa cells in a dose-dependent manner.
在这项研究中，通过反转录 PCR 克隆了来自日本河豚（Takifugu rubripes）的肿瘤坏死因子相关凋亡诱导配体（TRAIL）的 cDNA。TRAIL 的开放阅读框由 870 个碱基组成。TRAIL 的推导氨基酸序列与其他硬骨鱼的序列具有高度同源性。重组可溶性 TRAIL 与一个小泛素相关修饰物基因融合，以提高在大肠杆菌 BL21（DE3）中的可溶性表达水平。体外实验中，3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测表明，纯化的可溶性 TRAIL 能够以剂量依赖的方式诱导 Jurkat 和 HeLa 细胞凋亡。
