肿瘤坏死因子相关的凋亡诱导配体(TRAIL)cDNA的克隆与表达

肿瘤坏死因子相关凋亡诱导配体（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒｒｅｌａｔｅｄａｐｏｐｔｏｓｉｓｉｎｄｕｃｉｎｇｌｉｇａｎｄ，ＴＲＡＩＬ）是新近发现的肿瘤坏死因子家族成员，亦称凋亡素２配体（Ａｐｏ２ｌｉｇａｎｄ，Ａｐｏ２Ｌ）［１，２］．由于ＴＮＦ参与机体的免疫调节和炎症反应，并发挥细胞毒作用，因而为世人所瞩目［３，４］．研究表明，ＴＲＡＩＬ与ＴＮＦ和Ｆａｓ／Ａｐｏ１配体一样，为典型的Ⅱ型跨膜蛋白，Ｃ端细胞外区域形成同源三聚体的亚结构，而Ｎ端１５～４０氨基酸为疏水区域并形成跨膜结…

Cloning and Expression of Human TNF Related Apoptosis inducing Ligand in E. coli

Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.