Molecular analysis and expression of major outer membrane protein W of Vibrio alginolyticus isolated from flounder (Paralichthys olivaceus)
2010
Yang, D.K., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Kim, J.Y., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Kim, C.L., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Lee, K.W., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Jang, H., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Son, S.W., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Song, J.Y., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea
Vibrio alginolyticus (V. alginolyticus) is one of the vibriosis pathogens in human and several fish including flounder (Paralichthys olivaceus). Outer membrane proteins (Omp) of bacteria play an important role in the host immune response. In this study, the OmpW genes of 13 Korean V. alginolyticus isolates collected from flounders during a period between April and September of 2008 were subjected to molecular analysis. The similarity of the nucleotide sequences of the OmpW gene among all 13 field isolates in Korea ranged from 97.9 to 100%. The nucleotide analysis showed that one of the isolates. KBVV8. had a high similarity with that of V. alginolyticus (98.3%), V. harveyi (92.1%), V. parahaemolyticus (92.1%). but a low similarity with that of V. cholerae (69.1%). The KBVV8 isolate had 94.4 and 72,2% amino acid similarties with V. parahaemolyticus and V, cholera, respectively. Finally, we cloned an OmpW gene from V. alginolyticus (KBVV8 isolate) and expressed it in sf-9 cells using baculovirus expression system Western blot assay revealed that recombinant OmpW protein had immunoreaction with anti-six histidine monoclonal antibody corresponding to the predicted molecular weight of 23 kDa. The present study provides the scientific basis that recombinant OmpW protein can be used as a novel vaccine candidate against infection by V. alginolyticus in flounder.
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