Recombinant outer membrane protein A (OmpA) of Edwardsiella tarda, a potential vaccine candidate for fish, common carp |
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Authors: | Maiti Biswajit Shetty Mahesh Shekar Malathi Karunasagar Iddya Karunasagar Indrani |
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Institution: | UNESCO-MIRCEN for Marine Biotechnology, Department of Fishery Microbiology, Karnataka Veterinary, Animal & Fisheries Sciences University, College of Fisheries, Mangalore-575002, India |
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Abstract: | Outer membrane protein A (OmpA) is a component of the outer membrane of Edwardsiella tarda and is wildly distributed in Enterobacteriaceae family. The gene encoding the OmpA protein was cloned from E. tarda and expressed in Escherichia coli M15 cells. The recombinant OmpA protein containing His6 residues was estimated to have a molecular weight of ∼38 kDa. In Western blot the native protein showed expression at ∼36 kDa molecular weight which was within the range of major outer membrane proteins (36–44 kDa) observed in this study. All E. tarda isolates tested harbored the ompA gene and the antibody raised to this protein was seen to cross react with other Gram negative bacteria. The OmpA protein characterized in this study was observed to be highly immunogenic in both rabbit and fish. In Enzyme linked immunosorbent assay, rabbit antisera showed an antibody titer of 1: 128,000. Common carp vaccinated with recombinant OmpA protein elicited high antibody production and immunized fish showed a relative percentage survival of 54.3 on challenge. |
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Keywords: | Edwardsiella tarda OmpA Cloning and expression Western blot Vaccination Aquaculture ELISA |
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