重组SARS冠状病毒M蛋白的表达、纯化及鉴定  

Expression, Purification and Identification of Recombinant SARS Coronavirus Membrane Protein

作　　者：张晓丽[1] 王晶茹[1] 张燕[1] 陈茂林[1] 张伟[1] 杨晟[1] 姜卫红[1] 

机构地区：[1]中国科学院上海生命科学研究院植物生理生态研究所,上海200032

出　　处：《生物化学与生物物理学报》2003年第12期1140-1144,共5页

基　　金：中国科学院抗SARS科研基金;中国科学院上海生命科学研究院植物生理生态研究所所长基金资助项目~~

摘　　要：SARS冠状病毒是人的严重急性呼吸综合征的病原体。根据对其他种类冠状病毒的研究结果 ,膜蛋白 (M蛋白 )是病毒主要的结构蛋白 ,重组M蛋白可被用来作为抗原检测对应冠状病毒的感染和制备疫苗。SARS病毒M蛋白基因克隆到原核表达载体pMAL cRI中 ,利用N端和C端分别融合麦芽糖结合蛋白 (maltosebindingprotein和MxeGyrAinteinCBD的策略 ,在大肠杆菌中初步表达了重组M蛋白 ,并通过Western印迹和质谱对蛋白质进行了鉴定。重组蛋白质经亲和层析得到了部分纯化 ,纯化后的蛋白质将用于功能研究与诊断试剂盒的研制。A novel coronavirus (SARS-coronavirus, SARS-CoV) was discovered as the pathogen of the severe acute respiratory syndrome (SARS). According to studies with other coronaviruses, the membrane protein (M protein) is the main structural protein and the recombinant M protein may be useful as an antigen for detecting antibodies against coronavirus and for preparing vaccine. In this work, the M protein of SARS-CoV was expressed in E. coli as fusion protein with maltose binding protein at N-terminus and MxeGyrA intein CBD at C-terminus. The recombinant protein was identified by Western blot and mass spectrometry. The soluble parts of the cell crude extract were then partially purified by MBP affinity chromatography. The purified protein will be used for the studies on M protein's structure and the development of diagnostic method of SARS.

关 键 词：SARS 冠状病毒 重组M蛋白 表达 纯化 鉴定 严重急性呼吸综合征 基因组 

分 类 号：R346[医药卫生—基础医学]