SARS冠状病毒核衣壳(N)蛋白不同区域的原核表达  

Prokaryotic Expression of Different Regions of SARS-CoV Nucleocapsid Protein

作　　者：林鉴[1] 葛胜祥[1] 王颖彬[1] 罗文新[1] 吴婷[1] 李少伟[1] 程通[1] 张军[1] 夏宁邵[1] 

机构地区：[1]福建省医学分子病毒学研究中心

出　　处：《病毒学报》2005年第2期81-87,共7页Chinese Journal of Virology

基　　金：教育部跨世纪优秀人才培养计划(2002)

摘　　要：利用大肠杆菌表达系统对SARS冠状病毒的核衣壳(N)蛋白全长及N末端或/和C末端缺失突变体进行了表达,共表达了39个重组蛋白,表达量在15%～30%之间。分别利用电洗脱或金属鳌合介质纯化重组蛋白,用蛋白印迹实验检测纯化蛋白对SARS病人恢复期血清的反应性,结果发现全长N蛋白活性最好,其余的末端缺失蛋白均无法达到同一活性水平。由此说明N蛋白的完整性对于其优势表位的充分暴露是必要的。Nucleocapsid(N)protein of the severe acute respiratory syndrome associated coronavirus(SARS-CoV) , and its N or/and C terminal truncated mutants were expressed in E. coli .The yield levels of these thirty-nine recombinant proteins were from 15% to 30% .Recombinant proteins were purified by metal chelated affinity chromatography or by electro-elution. Western blot was used to detect the reactivity of these proteins with the convalescent sera of SARS patients. The results showed that the reactivity of the full length N protein is the best,which suggests that the integrity of N protein is important for exposure of the prominent epitopes.

关 键 词：重组蛋白 SARS冠状病毒 SARS病人 N蛋白 表位 实验检测 恢复期 核衣壳 利用 缺失突变体 

分 类 号：Q786[生物学—分子生物学] R373[医药卫生—病原生物学]