肿瘤坏死因子相关的凋亡诱导配体(TRAIL)可溶性胞外区在大肠杆菌中的高效表达

High Level Expression of Human TNF-Related Apoptosis-Inducing Ligand Extracellular Domain in E.Coli

肿瘤坏死相关凋亡诱导配体 (TNF relatedapoptosis inducingligand,TRAIL) ,也称为凋亡素 2配体(Apoptosisin2Ligand ,Apo2L) ,是肿瘤坏死因子超家族成员 ,能够激发肿瘤细胞选择性地发生凋亡 ,并对正常组织细胞几乎无毒性。TRAIL通过细胞表达受体 ,即死亡受体 4和 5 (DeathReceptor 4 ,DR4andDeathReceptor5 )发挥作用 ,而大多数肿瘤细胞表面表达这 2种受体。为了深入研究TRAIL在肿瘤治疗中的价值 ,我们采用RT PCR方法克隆了TRAIL基因 ,并对其细胞外可溶性区域 (1 1 4 2 81氨基酸残基 )在大肠杆菌中进行了表达。重组的TRAIL可溶性蛋白占细菌总蛋白的 4 5 %以上 。

TNF-related apoptosis-inducing ligand (TRAIL , also called Apo2L) can trigger tu mo r cell apoptosis.As TRAIL mRNA is expressed in many tissues, suggesting that i t might be nontoxic to normal cells.Many recent research works show that TRAIL c an induce cell death by death receptor 4 and 5, which are mostly expressed in ma ny tumor cells.To investigate TRAIL's therapeutic potential, we generated in E . coli a very potently active solube version of the native human TRAIL protein. T he recombinant protein was highly expressed in E.coli BL21 up to 30% of ba cteri al protein.The recombinant protein was tested in several tumor cells for its a ctivity