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Virologica Sinica, 20 (3) : 217-220, 2005
Research Article
Purification of Glutathione S-transferase Fusion Proteins of SARS Coronavirus Spike Protein fragment 2 and Preparation of Anti-GST-S2 Polyclonal Antibody
 Correspondence:
(200.99KB)  
Abstract
To study the immunogenicity of spike protein of SARS coronavirus, an expressional plasmid was constructed encoding partial sequence of SARS Coronavirus spike protein from 2170 bp to 2814bp, named fragment-2(S2). The gene sequence encoding S2 was amplified by RT-PCR from SARS Coronavirus genome RNA, cloned to pMD18-T vector and subcloned to pGEX-4T-2. E.coli JM109 that contained recombinant expression vector pGEX-S2 was amplified and fusion protein expression was induced by 1mmol/L IPTG at 37℃. Protein was extracted and purified by GSH-Sepharose affinity chromatography. After identified by the serum of SARS patient by Western-blot, the purified protein GST-S2 was used to immunize NIH mice three times at two weeks interval, the immunized mice produced high titer anti-GST-S2 polyclonal antibody. It lies a basis for the future study of subunit vaccine of SARS coronavirus.
  Published online: 20 Jun 2005
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