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.
出处
《病毒学报》
CAS
CSCD
北大核心
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))
