title: Preparation and characterization of polyclonal antibody against severe acute respiratory syndrome-associated coronavirus spike protein.
abstract: 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.
authors.affiliation: Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Ren
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics; D007544:Isopropyl Thiogalactoside; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011817:Rabbits; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics*; D064370:Spike Glycoprotein, Coronavirus; D014170:Transformation, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D017931:DNA Primers; D008562:Membrane Glycoproteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus; D007544:Isopropyl Thiogalactoside