Analysis of interferon gamma protein expression in zebrafish (Danio rerio)

Sohye Yoon, Ayham Alnabulsi, Steven Bird, Jun Zou, Christopher John Secombes

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)79-86
Number of pages8
JournalFish & Shellfish Immunology
Volume79-86
Issue number57
Early online date15 Aug 2016
DOIs
Publication statusPublished - Oct 2016

Keywords

  • Zebrafish
  • IFN-γ
  • Monoclonal antibody
  • Protein expression
  • PHA
  • Antigen re-stimulation

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