title: Vaccinia virus expression and sequence of an avian influenza nucleoprotein gene: potential use in diagnosis.
abstract: The nucleoprotein (NP) gene from avian influenza strain A/Shearwater/Aust/1/72 (H6N5) was cloned, sequenced, and expressed in vaccinia virus for the production of potent sera in immunised rabbits. The NP gene is 1565 bp and shares greater than 95% amino acid sequence identity with other NPs of the avian subtype. The recombinant NP expressed by vaccinia virus comigrated with endogenous A/Shearwater/Aust/1/72 NP by Western blot analysis. Polyclonal rabbit sera raised against recombinant NP was evaluated in an antigen capture ELISA system as a potential diagnostic tool for the detection of avian influenza. All type A strains, comprising several HA and NA subtypes, but not type B nor other avian viruses, were detected.
authors.affiliation: St. Vincent's Institute of Medical Research, Fitzroy, Australia.
authors.forename: V R
authors.initials: VR
authors.lastname: Harley
authors.forename: P J
authors.initials: PJ
authors.lastname: Hudson
authors.forename: B E
authors.initials: BE
authors.lastname: Coupar
authors.forename: P W
authors.initials: PW
authors.lastname: Selleck
authors.forename: H
authors.initials: H
authors.lastname: Westbury
authors.forename: D B
authors.initials: DB
authors.lastname: Boyle
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001483:Base Sequence; D001717:Birds; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004279:DNA, Viral; D004797:Enzyme-Linked Immunosorbent Assay; D005814:Genes, Viral; D009980:Influenza A virus / Q000235:genetics* / Q000276:immunology; D005585:Influenza in Birds / Q000175:diagnosis*; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000235:genetics* / Q000276:immunology; D011237:Predictive Value of Tests; D016601:RNA-Binding Proteins; D013937:Thymidine Kinase / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics* / Q000276:immunology; D014758:Viral Core Proteins; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D004279:DNA, Viral; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins; D014764:Viral Proteins; D013937:Thymidine Kinase