Expression of recombinant human IFN-γ protein in soybean (Glycine max L.)

Abstract

In recent years, the diagnostic and therapeutic application of recombinant proteins is increasing rapidly. One of these proteins is human interferon-γ (hIFN-γ), which shows great value in therapy against the viruses. The different biological drugs, such as cytokines, vaccines, growth factors, and antibodies are produced extensively through plant-based expression systems. In the present investigation, we report the successful expression of recombinant human interferon-γ (hIFN-γ) protein in soybean by Agrobacterium-mediated. For this purpose, the recombinant construct (pCAMBIA1304-hIFN-γ) was transferred in the GV3101 strain of Agrobacterium tumefaciens and applied for stable transformation of soybean explants. The results have confirmed the presence of the hIFN-γ gene in transgenic soybean by PCR, and also, the integration and copy number of the hIFN-γ gene was confirmed by southern blot analysis. The expression of the hIFN-γ transcription was confirmed by RT-PCR. Moreover, the total protein of transgenic soybean was extracted, and the expression of the hIFN-γ proteins was determined by western blot. Finally, bioactivity assay demonstrates that recombinant hIFN-γ protein was active biologically and protects the Vero cells versus infection that produced by the VSV. This investigation can be used as a basis for further researches in the large scale production of hIFN-γ and other recombinant proteins in soybean. For the first time, the successful transformation and expression of human gamma interferon gene in soybean plant by Agrobacterium tumefaciens were reported and the antiviral activity of recombinant hIFN-γ protein produced in soybean was confirmed.