【摘要】 将含有人TRAIL蛋白表达质粒的大肠杆菌以IPTG诱导表达 ,表达蛋白主要以包涵体形式存在 .经菌体破碎、包涵体分离、抽提及复性 ,用Ni NTASuperflow柱层析及进一步的离子交换色谱分离纯化获得纯蛋白 .SDS PAGE显示纯化的蛋白为 19kD的单一条带 .Westernblot鉴定表明 ,纯化的表达产物与抗人TRAIL蛋白的抗体发生特异性免疫反应 .抗肿瘤生物活性测定表明该纯化蛋白对肿瘤细胞的体外生长具有显著的抑制作用 .更多还原

【Abstract】 E.coli BL21 baring a expression plasmid of human TRAIL protein was induced for expression with IPTG.The protein expressed in a form of inclusion bodies and was purified through cell disruption, inclusion bodies extraction, solubilization renaturation, and a combination of Ni-NTA Superflow affinity and ion-exchange chromatography. The result of SDS-PAGE and Western blot demonstrated that the purified protein was a single band with molecular weight of 19kD and reacted specially with anti-human TRIAL IgG. Cytotoxic activity assay showed that the purified TRAIL protein had good antitumor activity in vitro.更多还原