Sequence and expression analysis of an interferon stimulated gene (ISG15) from Atlantic cod ( Gadus morhua L.)

• Published in: Developmental & Comparative Immunology Vol. 31; no. 2; pp. 156 - 171
• Main Authors: Seppola, Marit, Stenvik, Jørgen, Steiro, Kari, Solstad, Terese, Robertsen, Børre, Jensen, Ingvill
• Format: Journal Article
• Language: English
• Published: United States Elsevier Ltd 2007
• Subjects: Amino Acid Sequence; Animals; Atlantic cod; Base Sequence; Brackish; Cloning, Molecular; Cytokines - chemistry; Cytokines - genetics; Danio rerio; Fish; Gadus morhua; Gadus morhua - immunology; Immunity, Innate; Innate immunity; Interferons - pharmacology; Intron; Introns; ISG15; ISG15 promoter; Marine; Molecular Sequence Data; Phylogeny; Poly I:C; Promoter Regions, Genetic; Real time PCR; Teleostei; Ubiquitins - chemistry; Ubiquitins - genetics; Vibrio; A, Atlantic; ORF; EST; Atlantic cod; Innate immunity; SSH; ISG15; i.p; Poly I:C; LPS; IFN; UTR; Intron; SNP; GAS; Fish; ISGylation; Real time PCR; ISG; IRF; ISG15 promoter; UBL; ISRE
• ISSN: 0145-305X; 1879-0089; 1365-2567
• DOI: 10.1016/j.dci.2006.05.009

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.