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

作者：Yoon Sohye^*; Alnabulsi Ayham; Wang Ting Yu; Lee Po Tsang; Chen Tzong Yueh; Bird Steve; Zou Jun; Secombes Christopher J^*

来源：Fish & Shellfish Immunology, 2016, 57: 79-86.

DOI：10.1016/j.fsi.2016.08.023

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

IFN-gamma 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-gamma protein expression and bioactivity in fish. Hence, the current study focused on developing a monoclonal antibody (moAb) against zfiFN-gamma. We show that the zfIFN-gamma moAb specifically recognises E. coli produced recombinant IFN-gamma protein and zfIFN-gamma 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-gamma protein is produced as a dimer, and that a good correlation exists between transcript expression levels and protein levels.

• 出版日期2016-10