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ThesisItem Open Access Development of recombinant protein vaccine against Vibrio infection of aquatic animals(KARNATAKA VETERINARY ANIMAL AND FISHERIES SCIENCE'SUNIVERSITY, BIDAR-585226, 2011-09) Mohammed Abdullatif Hamod; I KARUNASAGSR; S.K.KOTE; M. N. VENUGOPAL; B A SAMSUNDAR; M.M.MARGALVibrio anguillarum is a naturally flora of aquatic environment and the most common pathogen of marine and brackish water finfish culture. Use of anti-microbial agents to control the diseases has ceased to be the method of choice since it results in drug resistance, accumulate drug residues and environmental imbalance problem. Therefore development of vaccines especially recombinant protein vaccine is a recent strategy to address diseases prevention. Outer membrane proteins (OMPs) are highly immunogenic bacterial components due to their exposed epitopes on the cell surface. In this study nine isolates of V. anguillarum were obtained, identified and characterized by conventional biochemical tests and molecular method. The ompU, ompK and OmpV genes of V. anguillarum were amplified by PCR, cloned and sequenced. Bioinformatics analysis of the structures of targeted OMPs and immunogenic potency evaluation were carried out to facilitate the development of recombinant vaccine for finfish. OmpU (35 kDa), OmpK (28 kDa), and OmpV (27 kDa) were cloned, over expressed and polyclonal antibodies were raised in rabbit and they were identified by ELISA and western blotting. Challenge studies revealed that OmpK and OmpV were highly effective in eliciting immune respond and affording protection. Thus they are considered potential candidates for subunit vaccine to protect fish against V. anguillarum disease. The possible function of ompK gene in V. anguillarum was studied. Factors affecting ompK gene expression in V. anguillarum suggests its role in iron uptake and resistance to bile salts.