title: Baculovirus expression of beak and feather disease virus (BFDV) capsid protein capable of self-assembly and haemagglutination.
abstract: Beak and feather disease virus (BFDV) is a common avian circovirus infection of wild Psittaciformes and is a recognised threat to endangered psittacine species. Currently, there is a requirement to develop BFDV antigen for diagnostic purposes and since efforts to propagate BFDV in vitro have so far been unsuccessful the entire coding region of BFDV ORF C1 was expressed in Sf9 insect cells using a baculovirus expression system. The entire coding region of BFDV ORF C1, the presumptive capsid, was expressed in Sf9 insect cells using baculovirus expression system. Electron microscopic examination of negatively stained material demonstrated that the recombinant protein self-assembled to produce virus-like particles (VLPs) thus confirming that ORF C1 is likely to be the sole determinant for capsid construction in vivo. BFDV VLPs also possessed haemagglutinating activity which provides further evidence that self-assembled BFDV VLPs retain receptor mediated biological activity and that the determinants for BFDV haemagglutination activity rely solely on the capsid protein. The recombinant protein reacted with anti-BFDV sera from naturally immune parrots and cockatoo and from chickens experimentally inoculated with native BFDV in both Western blots and haemagglutination inhibition (HI) assay. BFDV VLPs were also a suitable replacement antigen for serological detection of BFDV antibody by HI.
authors.affiliation: School of Veterinary and Biomedical Science, Murdoch University, South St., Murdoch, 6150 WA, Australia.
authors.forename: Meredith E
authors.initials: ME
authors.lastname: Stewart
authors.forename: Nicolai
authors.initials: N
authors.lastname: Bonne
authors.forename: Patrick
authors.initials: P
authors.lastname: Shearer
authors.forename: Bahman
authors.initials: B
authors.lastname: Khalesi
authors.forename: Margaret
authors.initials: M
authors.lastname: Sharp
authors.forename: Shane
authors.initials: S
authors.lastname: Raidal
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D016367:Baculoviridae / Q000378:metabolism*; D001717:Birds; D015153:Blotting, Western / Q000379:methods; D002213:Capsid / Q000502:physiology; D036022:Capsid Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D018173:Circoviridae Infections / Q000097:blood / Q000175:diagnosis / Q000821:virology; D017925:Circovirus / Q000737:chemistry* / Q000502:physiology; D006385:Hemagglutination Inhibition Tests / Q000379:methods; D006387:Hemagglutination, Viral; D007106:Immune Sera / Q000276:immunology; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000096:biosynthesis; D019065:Virus Assembly
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D007106:Immune Sera; D011994:Recombinant Proteins