人TRAIL基因胞外段的化学合成和克隆

Chemical synthesis and Cloning of the Human TRAIL gene extracellular region

根据大肠杆菌高表达序列重新设计了人肿瘤坏死因子相关的凋亡诱导配体(TRAIL)胞外段的基因编码区的核苷酸序列,应用化学合成的方法合成了多个互补DNA片段,经互补片段分别退火和连接后,将完整编码区克隆于pMD18-T质粒中,进行DNA序列分析,获得了与设计序列完全一致的含有完整编码区的人TRAIL胞外段基因,并成功构建了高效的TRAIL胞膜外段原核表达载体pTWIN1/TRAIL_(111-281)。

The coding regions nucleotide sequence of the human TNF-related apoptosis-inducing ligand (TRAIL) gene extracellular region was designed according to high level expression sequences of Ecoli Several overlapping complementary oligo-nueleotide fragments were synthesized with chemical synthesis method By annealing and ligating the corresponding oligo-nucleotide, the fragment that cover the extracellular region of TRAIL gene were cloned in plasmid pMD-18T The clones of interest were identified by DNA sequencing The cloned extracellular region of TRAIL gene fragments were recombined in to plasmid pTWIN1 The results showed that the redesigned full length coding region of human extracellular region of TRAIL gene was obtained as expected.