作　　者： ; ; ; ; ; ; ;

机构地区： 南方医科大学

出　　处： 《病毒学报》 2005年第4期303-304,共2页

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： 冠状病毒 蛋白 昆虫细胞 严重急性呼吸综合征 纯化 表达 宿主细胞 相互作用 细胞受体

领　　域： [生物学] [农业科学] [农业科学] [农业科学]