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 ve...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.展开更多
文摘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.
