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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
【Abstract】 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.
【Key words】 <Keyword>SARS-CoV; spike protein; baculovirus;
- 【文献出处】 病毒学报 ,Chinese Journal of Virology , 编辑部邮箱 ,2005年04期
- 【分类号】R392
- 【被引频次】4
- 【下载频次】225