Cloning and identification of the infectious salmon anaemia virus haemagglutinin.
Infectious salmon anaemia virus (ISAV) is an orthomyxo-like virus that causes serious disease in Atlantic salmon (Salmo salar). Like the orthomyxoviruses, ISAV has been shown to possess haemagglutinin (HA) activity. This study presents the cloning, expression and identification of the ISAV HA gene, which was isolated from a cDNA library by immunoscreening. The HA gene contained an ISAV-specific conserved nucleotide motif in the 5' region and a 1167 bp open reading frame encoding a protein with a predicted molecular mass of 42.4 kDa. The HA gene was expressed in a baculovirus system. A monoclonal antibody (MAb) shown previously to be directed against the ISAV HA reacted with insect cells infected with recombinant baculovirus. Salmon erythrocytes also adsorbed to these cells and adsorption was inhibited by the addition of either the ISAV-specific MAb or a polyclonal rabbit serum prepared against purified virus, confirming the virus specificity of the reaction. Immunoblot analyses indicated that ISAV HA, in contrast to influenza virus HA, is not posttranslationally cleaved. Sequence comparisons of the HA gene from five Norwegian, one Scottish and one Canadian isolate revealed a highly polymorphic region that may be useful in epidemiological studies.
Krossøy
Bjørn
B
Department of Fisheries and Marine Biology, University of Bergen, Norway2.|Intervet Norbio, Thormøhlensgate 58, N-5008 Bergen, Norway1.
Devold
Magnus
M
Department of Fisheries and Marine Biology, University of Bergen, Norway2.
Sanders
Lisette
L
Intervet International BV, Wim de Körverstraat 35, 5831 Boxmeer, The Netherlands3.
Knappskog
Per Morten
PM
Department of Medical Genetics, Haukeland Hospital, Bergen, Norway4.
Aspehaug
Vidar
V
National Veterinary Institute, Oslo, Norway5.
Falk
Knut
K
National Veterinary Institute, Oslo, Norway5.
Nylund
Are
A
Department of Fisheries and Marine Biology, University of Bergen, Norway2.
Koumans
Sjo
S
Intervet International BV, Wim de Körverstraat 35, 5831 Boxmeer, The Netherlands3.
Endresen
Curt
C
Department of Fisheries and Marine Biology, University of Bergen, Norway2.
Biering
Eirik
E
Intervet Norbio, Thormøhlensgate 58, N-5008 Bergen, Norway1.
D000595:Amino Acid Sequence; D000740:Anemia / Q000662:veterinary*; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D005393:Fish Diseases / Q000821:virology*; D005399:Fishes; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D006389:Hemagglutinins, Viral / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000235:genetics* / Q000276:immunology; D009976:Orthomyxoviridae Infections / Q000662:veterinary*; D011994:Recombinant Proteins / Q000096:biosynthesis; D020317:Salmo salar; D016415:Sequence Alignment
D006389:Hemagglutinins, Viral; D011994:Recombinant Proteins