title: Sequence and expression analysis of an interferon stimulated gene (ISG15) from Atlantic cod (Gadus morhua L.).
abstract: Suppression subtractive hybridization was used to examine gene expression in Atlantic cod head kidney cells treated with polyinosinic polycytidylic acid (poly I:C). One of the most abundant genes was ISG15, showing 24-53% amino acid similarity to ISG15 from both mammals and teleosts. The promoter was cloned by genome walking and three potential interferon-stimulated response elements (ISREs) were identified. Analysis of the gene structure revealed a single intron in the 5' untranslated region (UTR) of cod ISG15, which also seems to be present in zebrafish and pufferfish ISG15. A quantitative real time PCR assay was established to monitor the gene expression of cod ISG15. Injection of cod with poly I:C strongly induced the expression of ISG15 in all organs investigated. Stimulation was most pronounced the first day with a gradual decline the following days. The expression of ISG15 in head kidney cells was also induced in vitro by treatment with poly I:C, but not significantly with LPS. However, injection of formalin killed Vibrio anguillarum-induced ISG15 expression in head kidney.
authors.affiliation: Fiskeriforskning, Norwegian Institute of Fisheries and Aquaculture Research, Box 6122, N-9291 Tromsø, Norway. marit.seppola@fiskeriforskning.no
authors.forename: Marit
authors.initials: M
authors.lastname: Seppola
authors.forename: Jørgen
authors.initials: J
authors.lastname: Stenvik
authors.forename: Kari
authors.initials: K
authors.lastname: Steiro
authors.forename: Terese
authors.initials: T
authors.lastname: Solstad
authors.forename: Børre
authors.initials: B
authors.lastname: Robertsen
authors.forename: Ingvill
authors.initials: I
authors.lastname: Jensen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D016207:Cytokines / Q000737:chemistry / Q000235:genetics*; D048389:Gadus morhua / Q000276:immunology*; D007113:Immunity, Innate; D007372:Interferons / Q000494:pharmacology*; D007438:Introns; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D014452:Ubiquitins / Q000737:chemistry / Q000235:genetics*
chemical_list: D016207:Cytokines; D014452:Ubiquitins; D007372:Interferons