In Silico Designing and Characterization of Outer Membrane Protein (ompc) Gene from Salmonella Enterica and Its Expression in Nicotiana Tabacum for Developing a Plant-based Vaccine Against Salmonellosis

Salmonella, a gram-negative bacteria, is the leading cause of foodborne illness globally. Two serovars of Salmonella, S. enteritidis and S. typhimurium are responsible for the majority of human salmonellosis. Prolonged salmonellosis caused by Salmonella species leads to the development of colon cancer, which is 3rd most common cancer in the world. Porins in the outer membrane of Salmonella can be used to elicit immune response. The production of plant-based vaccine against salmonellosis and the subsequent colon cancer using outer membrane proteins can be helpful for the people of developing countries. In this study, OmpC protein from Salmonella enteritidis was subjected to various bioinformatics tools which exhibited OmpC vaccine construct to be sufficiently immunogenic, non-allergenic, non-toxic and non-homologous to human proteins. Docking analysis showed strong interaction of OmpC vaccine model with TLR-4. After in silico analysis, this vaccine construct was expressed in tobacco plants via Agrobacterium-mediated transformation. Gateway (R) cloning was used to clone OmpC gene. Transformation and integration of transgene within tobacco plants was confirmed through conventional PCR. qRT-PCR was done for expression analysis and copy number calculated was 2. The expressed OmpC protein accumulated up to 0.42% of total soluble protein. Immunization of mice with total soluble protein (TSP) and purified OmpC protein generated significant level of anti-OmpC antibodies. The vaccine candidate also demonstrated significant protective effect in mice upon challenging with Salmonella typhimurium. To the best of our knowledge, this is the first study reporting the expression of OmpC antigen in plants for potential use as vaccine against salmonellosis.