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SARS冠状病毒S蛋白在昆虫细胞中的表达和纯化

Expression of Spike Protein of SARS Coronavirus in a Baculovirus System and Its Purification

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【作者】 李志杰要国华刘靖华赵善超邓鹏徐军姜勇

【Author】 LI Zhi-jie~1, YAO Guo-hua~2, LIU Jing-hua~1, ZHAO Shan-chao~1, DENG Peng~1, XU Jun~2, JIANG Yong~(1*)(1. Key Laboratory of Functional Proteomics of Guangdong Province, Nanfang Medical University, Guangzhou510515, China;2. Guangzhou Respiratory Diseases Institute, Guangzhou Medical College, Guangzhou510120, China)

【机构】 南方医科大学广东省功能蛋白质组学重点实验室广州医学院广州呼吸病研究所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510120广州510515广州510515

【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 pFastBacTM 1, forming a recombinant plasmid pFastBacTM 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.

【关键词】 <Keyword>SARS-CoVspike proteinbaculovirus
【Key words】 <Keyword>SARS-CoVspike proteinbaculovirus
【基金】 国家自然科学基金(No.30340029);973SARS专项课题(No.2003CB514117);中德SARS合作研究项目(No.GZ238(202/11))
  • 【文献出处】 病毒学报 ,Chinese Journal of Virology , 编辑部邮箱 ,2005年04期
  • 【分类号】R392
  • 【被引频次】4
  • 【下载频次】225
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