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| 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 | |
| 引用本文： | 易艳萍 李楚芳 石玉岭 李林海 李平 黄维 王升启 马清钧 曹诚. 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[J]. 生物工程学报, 2003, 19(4): 392-396 |
| 作者姓名： | 易艳萍 李楚芳 石玉岭 李林海 李平 黄维 王升启 马清钧 曹诚 |
| 作者单位： | 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 |
| 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pET22b和pBV222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，可用于抗SARS抗体检测及亚单位疫苗研究。 |
| 关 键 词： | SARS病毒 核衣壳蛋白 膜蛋白 表达 |
| 文章编号： | 1000-3061(2003)04-0392-05 |
| 修稿时间： | 2003-05-22 |
Over-expression in Escherichia coli and Purification of Nucleocaspid and Membrane Protein of SARS Coronavirus |
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| YI Yan-Ping LI Chu-Fang SHI Yu-Ling LI Lin-Hai LI Ping HUANG Wei WANG Sheng-Qi MA Qing-Jun CAO Cheng. Over-expression in Escherichia coli and Purification of Nucleocaspid and Membrane Protein of SARS Coronavirus[J]. Chinese journal of biotechnology, 2003, 19(4): 392-396 | |
| Authors: | YI Yan-Ping LI Chu-Fang SHI Yu-Ling LI Lin-Hai LI Ping HUANG Wei WANG Sheng-Qi MA Qing-Jun CAO Cheng |
| Affiliation: | Beijing Institute of Biotechnology, Beijing 100850, China. |
| Abstract: | 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. |
| Keywords: | SARS coronavirus nucleocaspid protein membrane protein expression |
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