| SARS病毒BJ01株核壳蛋白在大肠杆菌中的表达与鉴定 |
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| 引用本文： | 陈水平,刘素辉,范宝昌,赵慧,姜涛,秦成峰,杨保安,秦鄂德.SARS病毒BJ01株核壳蛋白在大肠杆菌中的表达与鉴定[J].微生物学免疫学进展,2005,33(2):32-35. |
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| 作者姓名： | 陈水平 刘素辉 范宝昌 赵慧 姜涛 秦成峰 杨保安 秦鄂德 |
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| 作者单位： | 1. 军事医学科学院微生物流行病研究所,北京,100071
2. 新疆医科大学病原学教研室,新疆,830054 |
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| 摘 要： | 通过RTPCR从SARS病毒BJ01株的RNA中扩增全长N基因,经凝胶回收后插入pBAD/TOPOThioFusion表达载体。然后在Top10中利用阿拉伯糖进行诱导表达,在优化的条件下,表达产物以可溶性为主,表达量约占细菌可溶性总蛋白的47%。表达产物采用ProBond蛋白纯化系统纯化后,目的蛋白约占67%。经免疫印迹法检测,表达产物与SARS病人恢复期血清和兔的抗血清均可进行特异反应。BJ01株核壳蛋白在大肠杆菌中的高效可溶性表达为后续的ELISA试剂盒的研制奠定了基础。
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| 关 键 词： | SARS冠状病毒 核壳蛋白 可溶性表达 纯化 |
| 文章编号： | 1005-5673(2005)02-0032-04 |
| 修稿时间： | 2004年11月3日 |
Expression and detection of nucleocapsid protein of SARS BJ01 strain in E.coli |
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| CHEN Shui-ping.Expression and detection of nucleocapsid protein of SARS BJ01 strain in E.coli[J].Progress In Microbiology and Immunology,2005,33(2):32-35. |
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| Authors: | CHEN Shui-ping |
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| Institution: | CHEN Shui-ping~ |
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| Abstract: | The full-length nucleocapsid gene of SARS BJ01 strain was amplified from SARS-CoV RNA with RT-PCR. Amplified products were purified from gel performance, then cloned into pBAD/TOPO ThioFusion vector. The expression was induced in E.coli Top10 strain with arabinose. Under optimized conditions, the expression proteins which were mainly soluble were approximate 47% of total soluble proteins of Top10. After purified with ProBond purification system, the expected proteins were about 67%. And the expressed products could specifically react with convalescent sera of patients and antisera of rabbit against SARS. It laid a foundation for development of ELISA kit in detection SARS by using the highly expressed soluble products of nucleocapsid protein originated from BJ01 strain. |
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| Keywords: | SARS-CoV Nucleocapsid protein Soluble expression Purification |
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