title: Functional analysis of an orange-spotted grouper (Epinephelus coioides) interferon gene and characterisation of its expression in response to nodavirus infection.
abstract: We cloned and sequenced 2C I-IFN, a two-cysteine containing type I interferon (I-IFN) gene, in orange-spotted grouper (Epinephelus coioides). The cDNA has 769 base pairs, the protein has 172 amino acids, and the predicted signal peptide has 18 amino acids with two cysteines. This gene is similar to I-FNs from sea bass and other teleosts. 2C I-IFN has 5 exons and 4 introns, also similar to other teleost I-IFNs. Immunohistochemical (IHC) analysis indicated that expression is predominantly membrane-localized in healthy grouper, but has a zonal distribution in nodavirus-infected grouper. Grouper infected with nodavirus had elevated levels of 2C I-IFN at 72 h and Mx at days 6-7. Recombinant 2C I-IFN activated grouper Mx, leading to upregulated antiviral activity. The grouper Mx promoter was highly induced after treatment with recombinant 2C I-IFN. The present results suggest that expression of grouper 2C I-IFN may participate in the immunologic barrier function against nodavirus.
authors.affiliation: Laboratory of Molecular Genetics, Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Translational Center for Marine Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Agriculture Biotechnology Research Center, National Cheng Kung University, Tainan 70101, Taiwan.
authors.forename: Young-Mao
authors.initials: YM
authors.lastname: Chen
authors.affiliation: Department of Nursing, Tzu Hui Institute of Technology, Pingtung 92641, Taiwan.
authors.forename: Cham-En
authors.initials: CE
authors.lastname: Kuo
authors.affiliation: Laboratory of Molecular Genetics, Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Translational Center for Marine Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan.
authors.forename: Guan-Ru
authors.initials: GR
authors.lastname: Chen
authors.affiliation: Laboratory of Molecular Genetics, Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Translational Center for Marine Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Agriculture Biotechnology Research Center, National Cheng Kung University, Tainan 70101, Taiwan.
authors.forename: Yu-Ting
authors.initials: YT
authors.lastname: Kao
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Secombes
authors.affiliation: Laboratory of Molecular Genetics, Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Translational Center for Marine Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Agriculture Biotechnology Research Center, National Cheng Kung University, Tainan 70101, Taiwan; University Center for Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan; Research Center of Ocean Environment and Technology, National Cheng Kung University, Tainan 70101, Taiwan. Electronic address: ibcty@mail.ncku.edu.tw.
authors.forename: Tzong-Yueh
authors.initials: TY
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001492:Bass / Q000235:genetics* / Q000276:immunology / Q000821:virology; D002460:Cell Line; D005393:Fish Diseases / Q000235:genetics / Q000276:immunology* / Q000821:virology; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000276:immunology; D007113:Immunity, Innate; D007372:Interferons / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins / Q000235:genetics / Q000378:metabolism; D030381:Nodaviridae / Q000276:immunology*; D009928:Organ Specificity; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D012327:RNA Virus Infections / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D017422:Sequence Analysis, DNA
chemical_list: D029941:Fish Proteins; D064547:Myxovirus Resistance Proteins; D007372:Interferons
keywords: Antiviral protein; Mx; Nodavirus; Two-cysteine containing type I interferon (2C I-IFN) gene