重组人源脑红蛋白的高效表达、纯化及鉴定

脑红蛋白(Neuroglobin，NGB)是新发现的主要表达于脊椎动物神经元及视网膜中的第三类携氧珠蛋白。已有诸多报道认为脑红蛋白在缺氧缺血性脑损伤的神经保护过程中，作为活性氧簇(ROS)的清除剂或缺氧信号的感受器起重要作用，但其神经保护作用的具体机制不明。显然，实现该蛋白的制备对于研究其功能具有重要作用。基于此，通过RT-PCR从人胎脑中扩增出脑红蛋白cDNA并克隆到原核表达载体pBV220，通过测序鉴定正确后转化至大肠杆菌HB101中诱导表达，表达产物超声破碎后经凝胶过滤柱Sephacryl S-200和阴离子交换柱Q Sepharose FF纯化，最后通过Sephadex G-25脱盐。经15% SDS-PAGE和Western blot检测及质谱和蛋白序列分析鉴定制备的蛋白样品为脑红蛋白。本文采用两步法实现了脑红蛋白高效特异性纯化，为后期基于脑红蛋白神经保护作用的功能及机制研究奠定了重要基础。

Expression,Purification and Identification of Recombinant Human Neuroglobin

Neuroglobin(NGB),widely and specially expressed in neurons of various vertebrates,was reported to be a scavenger of reactive oxygen species and/or a stress-responsive sensor for neuroprotection of hypoxic and ischemic insults.However,the underlying mechanism remained unknown.It is important for the functional study of NGB by preparing the protein samples.To address it,the human neuroglobin cDNA fragment was amplified by RT-PCR from human fetus brain and cloned into the prokaryotic expression vector pBV220,a...