SARS冠状病毒S蛋白在昆虫细胞中的表达和纯化

导致严重急性呼吸综合征(sevcre acute rcspiratory syndrome，SARS)的元凶是一种新型的冠状病毒(SARS coronavirus，SARS-CoV)。SARS-CoV感染入侵宿主细胞关键的一环是病毒自身的棘突蛋白(spike protein，S-protein)与细胞受体的相互作用，故而S蛋白己成为SARS研究的主要热点。

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

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.