📄 Preparation and characterization of polyclonal antibody against severe acute respiratory syndrome-associated coronavirus spike protein

A truncated gene (designated S1) encoding the receptor-binding domain (RBD) in the spike (S) protein of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) was amplified by PCR. The gene was cloned into prokaryotic expression vector pGEX-6P-1, resulting in a recombinant plasmid pGEX-SARS-S1. Subsequently, pGEX-SARS-S1 was transformed into host cells BL21(DE3)pLysS, and the expression of the S1 protein was induced by isopropyl β-D-thiogalactoside (IPTG). Polyclonal antibody against SARS-CoV S1 protein was generated in a rabbit immunized with the purified S1 protein. The reactivity of the antibody to the SARS-CoV S1 protein was confirmed by Western blot analysis. ELISA indicated that the antibody against SARS-CoV S1 protein had no cross reaction with S1 proteins of transmissible gastroenteritis virus, a porcine coronavirus, and infectious bronchitis virus, an avian coronavirus. The SARS-CoV S1 protein and its antibody are valuable reagents for related studies. © Copyright 2010, Mary Ann Liebert, Inc. 2010.

keywords 🔗 severe acute (1373) 🔗 bronchitis virus (233) 🔗 receptor-binding domain (99) 🔗 host cell (262) 🔗 respiratory syndrome-associated (90) 🔗 gastroenteritis virus (188) 🔗 infectious bronchitis (235) 🔗 respiratory syndrome (2004) 🔗 acute respiratory (1734) 🔗 syndrome-associated coronavirus (88) 🔗 transmissible gastroenteritis (226)

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