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

作　　者：李志杰[1] 要国华[2] 刘靖华[1] 赵善超[1] 邓鹏[1] 徐军[2] 姜勇[1] 

机构地区：[1]南方医科大学广东省功能蛋白质组学重点实验室,广州510515 [2]广州医学院广州呼吸病研究所,广州510120

出　　处：《病毒学报》2005年第4期303-304,共2页Chinese Journal of Virology

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))

摘　　要：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.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.

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 

分 类 号：Q939.47[生物学—微生物学] S852.61[农业科学—基础兽医学]