| SARS冠状病毒S蛋白在昆虫细胞中的表达和纯化 |
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| 引用本文： | 李志杰,要国华,刘靖华,赵善超,邓鹏,徐军,姜勇. SARS冠状病毒S蛋白在昆虫细胞中的表达和纯化[J]. 病毒学报, 2005, 21(4): 303-304 |
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| 作者姓名： | 李志杰 要国华 刘靖华 赵善超 邓鹏 徐军 姜勇 |
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| 作者单位： | 南方医科大学,广东省功能蛋白质组学重点实验室,广州,510515;广州医学院,广州呼吸病研究所,广州,510120 |
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| 基金项目： | 国家自然科学基金;30340029; |
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| 摘 要： | 导致严重急性呼吸综合征(sevcre acute rcspiratory syndrome，SARS)的元凶是一种新型的冠状病毒(SARS coronavirus，SARS-CoV)。SARS-CoV感染入侵宿主细胞关键的一环是病毒自身的棘突蛋白(spike protein，S-protein)与细胞受体的相互作用，故而S蛋白己成为SARS研究的主要热点。
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| 关 键 词： | SARS冠状病毒 S蛋白 昆虫细胞 严重急性呼吸综合征 syndrome SARS-CoV 纯化 表达 宿主细胞 相互作用 细胞受体 |
| 文章编号： | 1000-8721(2005)04-0304-02 |
Expression of Spike Protein of SARS Coronavirus in a Baculovirus System and Its Purification |
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| LI Zhi-jie,YAO Guo-hua,LIU Jing-hua,ZHAO Shan-chao,DENG Peng,XU Jun,JIANG Yong. Expression of Spike Protein of SARS Coronavirus in a Baculovirus System and Its Purification[J]. Chinese journal of virology, 2005, 21(4): 303-304 |
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| Authors: | LI Zhi-jie YAO Guo-hua LIU Jing-hua ZHAO Shan-chao DENG Peng XU Jun JIANG Yong |
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| Affiliation: | LI Zhi-jie~1,YAO Guo-hua~2,LIU Jing-hua~1,ZHAO Shan-chao~1,DENG Peng~1,XU Jun~2,JIANG Yong~ |
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| Abstract: | For expressing the recombinant S protein in spodoptera fragiperda(Sf9) cells, the full-length cDNA of S protein was firstly amplified from plasmid pET-14b/S by means of PCR and subcloned into baculovirus transfer vector pFastBac~(TM) 1, forming a recombinant plasmid pFastBac~(TM) 1/S. It was then transposited with baculovirus shuttle vector (Bacmid) in the Max efficiency DH10BAC component cells by homologous recombination to construct the expression vector Bacmid/S. The Bacmid/S, after identified by PCR, was used to transfect Sf9 cells to express the target protein. The expressed recombinant S protein was analyzed by Western blot with human anti-SARS-CoV antiserum. The results showed that the recombinant S protein had been expressed correctly and efficiently in insect Sf9 cells and was purified by Ni-NTA affinity chromatography. (Western) blot showed that recombinant S protein could be recognized by human anti-SARS-CoV antiserum. |
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| Keywords: | SARS-CoV spike protein baculovirus |
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