Expression, Purification and Characterization of Recombinant Interferon-γ of Takifugu rubripes in Pichia pastoris

Interferon gamma (IFN-gamma), a major cytokine, has broad-spectrum antiviral function, in addition to play a key role in the process of immune regulation. In this study, in order to explore the biological roles of recombinant IFN-gamma (rIFN-gamma), IFN-gamma was cloned from Takifugu rubripes and expressed in Pichia pastoris under the control of an electrical process. P. pastoris were screened by high concentration culture medium of Zeocin, producing the high-copy number P. pastoris induced by culture medium of BMGY/BMMY. We used mass spectrometry (MS) analysis to identify the rIFN-gamma protein. The results showed that the molecular weight of rIFN-gamma was 24 kDa by SDS-PAGE gel electrophoresis, the bioactivity of rIFN-gamma was 2018 U/mL and the concentration of the rIFN-gamma protein was 0.242 mg/mL after inducing for 96 h. Also, we identified the rIFN-gamma protein of T. rubripes. rIFN-gamma could stimulate WISH cell proliferation, which was associated with recombinant protein concentration. These results provide a basis for application of the rIFN-gamma protein as a vaccine candidate against pathogen infection in aquaculture. (C) 2019 Friends Science Publishers