title: Analysis of interferon gamma protein expression in zebrafish (Danio rerio).
abstract: IFN-γ is a major effector cytokine, produced to induce type I immune responses. It has been cloned in several fish species including zebrafish, however to date few studies have looked at IFN-γ protein expression and bioactivity in fish. Hence, the current study focused on developing a monoclonal antibody (moAb) against zfIFN-γ. We show that the zfIFN-γ moAb specifically recognises E. coli produced recombinant IFN-γ protein and zfIFN-γ produced in transfected HEK293 cells, by Western blot analysis. Next we analysed the production of the native protein following expression induced by PHA stimulation of leukocytes in vitro or antigen re-stimulation in vivo. We show the IFN-γ protein is produced as a dimer, and that a good correlation exists between transcript expression levels and protein levels.
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK. Electronic address: sohyeyoon@abdn.ac.uk.
authors.forename: Sohye
authors.initials: S
authors.lastname: Yoon
authors.affiliation: Vertebrate Antibodies Ltd, Aberdeen, UK.
authors.forename: Ayham
authors.initials: A
authors.lastname: Alnabulsi
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK.
authors.forename: Ting Yu
authors.initials: TY
authors.lastname: Wang
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK.
authors.forename: Po Tsang
authors.initials: PT
authors.lastname: Lee
authors.affiliation: Laboratory of Molecular Genetics, College of Bioscience and Biotechnology, National Cheng Kung University, Taiwan.
authors.forename: Tzong-Yueh
authors.initials: TY
authors.lastname: Chen
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK; Molecular Genetics, Department of Biological Sciences, University of Waikato, Hamilton, New Zealand.
authors.forename: Steve
authors.initials: S
authors.lastname: Bird
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK.
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK. Electronic address: c.secombes@abdn.ac.uk.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Secombes
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D007962:Leukocytes / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D016207:Cytokines; D011994:Recombinant Proteins; D007371:Interferon-gamma
keywords: Antigen re-stimulation; IFN-γ; Monoclonal antibody; PHA; Protein expression; Zebrafish