SARS病毒核衣壳蛋白、膜蛋白在大肠杆菌中的高效表达和纯化  

Over-expression in Escherichia coli and Purification of Nucleocaspid and Membrane Protein of SARS Coronavirus

作　　者：易艳萍[1] 李楚芳[1] 石玉岭[2] 李林海[2] 李平[1] 黄维[1] 王升启[3] 马清钧[1] 曹诚[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]广州军区总医院检验科,广州510010 [3]军事医学科学院放射医学研究所,北京100850

出　　处：《生物工程学报》2003年第4期392-396,共5页Chinese Journal of Biotechnology

摘　　要：通过反转录 PCR获得了SARS冠状病毒核衣壳蛋白 (N)和膜蛋白 (M)基因 ,其序列分析结果与加拿大多伦多株完全一致。将M基因和N基因克隆到大肠杆菌表达载体pET2 2b和pBV2 2 2上 ,并在大肠杆菌中以包涵体及可溶形式获得高效表达。通过离子交换、金属螯合层析纯化获得电泳纯制品。所获得的核衣壳蛋白具有良好的抗原性 。Genes encoding nucleocaspid (N) and membrane(M)protein of SARS coronavirus were obtained by RT-PCR and were cloned into expression vector pET22b and pBV222. DNA sequencing showed that the genes cloned from a patient in Beijing were identical to the gene sequences from reported Toronto strain. The genes were over-expressed in E.coli either as inclusion body or as soluble form. The recombinant proteins were purified by ion-exchange, or ion-exchange followed by metal chelate affinity chromatography. The recombinant N protein was demonstrated highly antigenic and could be employed as antigen to detect SARS antibodies in ELISA system for SARS diagnosis.

关 键 词：SARS病毒 核衣壳蛋白 膜蛋白 大肠杆菌 表达 纯化 

分 类 号：R346[医药卫生—基础医学]