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| SARS冠状病毒PUMC2株N蛋白cDNA的克隆、表达和纯化 | |
| 作者姓名： | Tan XY Fan Z Wang HJ Shi L Yin B Ni AP Qin C Zou K Shen Y Yuan JG Qiang BQ Peng XZ |
| 作者单位： | 1. 中国医学科学院,中国协和医科大学,基础医学研究所医学分子生物学国家重点实验室,北京,100005 2. Department of Clinical Laboratory, PUMC Hospital, CAMS and PUMC, Beijing 3. Institute of Experimental Animal, CAMS and PUMC, Beijing |
| 摘 要： | 目的 原核表达并纯化SARS冠状病毒N蛋白。方法 用RT-PCR技术克隆SARS冠状病毒PUMC2株N蛋白全长cDNA,cDNA经序列分析鉴定后,克隆到pET32a表达载体,转化E．coli BL21进行原核表达并纯化出SARS冠状病毒N蛋白。结果 实现了SARS冠状病毒N蛋白的表达及纯化。结论 用基因重组方法表达的SARS冠状病毒N蛋白可为其进一步功能研究提供条件。 |
| 关 键 词： | SARS冠状病毒 N蛋白 |
| 修稿时间： | 2003年8月7日 |
Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein |
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| Tan XY,Fan Z,Wang HJ,Shi L,Yin B,Ni AP,Qin C,Zou K,Shen Y,Yuan JG,Qiang BQ,Peng XZ.Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein[J].Acta Academiae Medicinae Sinicae,2003,25(5):504-507. | |
| Authors: | Tan Xin-yu Fan Zheng Wang Hua-jin Shi Lei Yin Bin Ni An-ping Qin Chuan Zou Ke Shen Yan Yuan Jian-gang Qiang Bo-qin Peng Xiao-zhong |
| Institution: | National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 10005, China. |
| Abstract: | OBJECTIVE: To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain. METHODS: According to the published SARS coronavirus genome sequences, the full length cDNA of N protein from SARS coronavirus PUMC2 strain was cloned by RT-PCR and the cDNA was cloned into the pET32a expression vector. The recombinant N protein was expressed in E. coli BL21 (DE3), and purified by Ni(2+)-NTA. RESULTS: Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained. CONCLUSIONS: The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein. |
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