Engineering and expression of SUMO-G fusion protein (small ubiquitin-like modifier domain and rabies glycoprotein (G)) in Nicotiana benthamiana plant

Rabies is an acute contagious infection of the central nervous system caused by the rabies virus, the tenth most common fatal infectious disease. The disease spreads through domestic and wild animals, killing an estimated 60,000 people worldwide each year. Rabies glycoprotein (G) plays an important role in viral pathogenesis and functions as a protective antigen. Although human and animal vaccines against rabies are currently available, they are expensive, laborious to manufacture, and have relatively poor immunogenicity. One of the protein post-translational modifications, SUMOylation has been reported to regulate protein subcellular localization, protein-DNA binding, protein-protein interactions, transcriptional regulation, DNA repair, and genome organization. The addition of a highly stable construct (such as SUMO) to the N-terminus of a partner protein increases efficiency by increasing stability. SUMO fused with proteins of interest significantly improves expression and promotes solubility by aiding in correct folding of proteins. The plant-based transient expression system is a promising technology for the production of various recombinant proteins, including vaccine antigens, therapeutic proteins, antibodies and industrial enzymes. These systems offer fast production timelines, low-cost input, highly scalable, high production capacity, and superior benefits over other expression systems that are free of mammalian pathogens. In this study, the Sumo protein belonging to Sumolation, one of the post-translational modifications of the protein, and the surface glycoprotein (GP) of the rabies virus were successfully produced in a short time in the plant transient expression system as fusion (SumoGP). The sumo-specific protease SENP1 has also been successfully produced in a plant transient expression system with high purity. Coexpressions of the SENP1 and SumoGP fusion protein were not clearly expressed in the plant transient expression system. Activation studies on SENP1 are ongoing. KEYWORDS: Rabies G protein, Sumolation, SUMO, Plant Transient Expression System, Nicotiana Benthamiana, Rabies