人肿瘤坏死因子真核表达探讨  

Reseach on the eukaryotic expression of human tumor necrosis factor

作　　者：王宇学[1] 陈晓[1] 徐锐锋[2] 

机构地区：[1]湖北中医学院医学检验与技术系,湖北武汉430064 [2]汉阳铁路中心医院检验科,湖北武汉430050

出　　处：《温州医学院学报》2006年第2期116-118,共3页Journal of Wenzhou Medical College

摘　　要：目的:研究人肿瘤坏死因子真核表达。方法:重组质粒(pORB2TNF-α)和线性杆状病毒(AcNPV)共转染昆虫细胞系(sf21),裂解转染细胞,ELISA方法蛋白定位,裂解物电泳评价杆状病毒重组体(AcNPV-pORB2TNF-α)表达蛋白;用6χHis标签纯化蛋白,纯化蛋白进行蛋白印迹(Westernblot)。结果:转染细胞表达蛋白为可溶蛋白;转染AcNPV-pORB2TNF-α细胞裂解物电泳见35kDXylE蛋白条带;纯化蛋白电泳可见17kD蛋白条带并且Westernblot呈阳性。结论:用构建人肿瘤坏死因子杆状病毒重组体,成功表达TNF-α蛋白。Objective： To study the eukaryotic expression of human tumor necrosis factor. Methods：Recombinant plasmid （pORB2TNF- α ） and linear Baculovirus （AcNPV） co-transfect insect cells（sf21）, the transfected cells were lysised by lysising buffer. The expression fusion protein of the AcNPV-pORB2TNF-α was located by ELISA method and assessed by SDS-PAGE; Total protein of the lysate was purified by the 6 χ His tag, and Western blot. Results：The expression fusion protein of transfection cells was found in upper solutions. On a lane there was a 35kD XylE gene strap in the lysate of cells transfected by AcNPV-pORB2TNF-α . The purified protein was on 17kD site of lane and positive for Western blot. Conclusion：The TNF-α protein is expressed succesfully with the Baculovirus recombinant of human tumor necrosis fractor.

关 键 词：肿瘤坏死因子 杆状病毒 蛋白质印迹 

分 类 号：R3[医药卫生—基础医学]