수산해양교육연구 (Journal of Fisheries and Marine Sciences Education)

  • 제27권5호
  • /
  • Pages.1508-1521
  • /
  • 2015
  • /
  • 1229-8999(pISSN)
  • /
  • 2288-2049(eISSN)
한국수산해양교육학회 (The Korean Society for Fisheries and Marine Sciences Education)
권호 표지
DOI QR코드

DOI QR Code

한국에서 분리된 Vibrio harveyi 외막단백질의 유전적 차이와 넙치(Paralichthys olivaceus)에 대한 OmpW의 면역원성 분석

Genetic Variations of Outer Membrane Protein Genes of Vibrio harveyi Isolated in Korea and Immunogenicity of OmpW in Olive Flounder, Paralichthys olivaceus

  • 투고 : 2015.09.24
  • 심사 : 2015.10.13
  • 발행 : 2015.10.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Vibrio harveyi is a pathogenic marine bacterium causing systemic symptoms resulting in mass mortalities in fishes and shrimps in aquaculture. Outer membrane proteins(OMPs) are related to the pathogenicity and thus good targets for diagnosis and vaccination for Gram negative bacteria. Recently vaccination strategies using the OMPs have been suggested to control vibriosis in several fish species. In this study, we have isolated V. harveyi from diseased marine fishes from different regions of Korea and investigated genetic variations of four OMP genes including OmpK, OmpU, OmpV and OmpW. Consequently, OmpK and U genes could be divided into 3 subgroups of type I, II, III and type A, B, C, respectively, without any correlation with geographical regions and species while OmpV and W were highly homologous. OmpW gene of V. harveyi FP4138 was fully sequenced and predicted the deduced amino acid sequence to form ${\beta}-barrel$ with hydrophobic channel. Indeed, the immunogenicity of recombinant OmpW produced in Escherichia coli was assessed by vaccinating flounder. As a result, the high antibody response with antibody titer of $4.2{\pm}0.7$ and protection with relative percent survival of 60% against artificial infection of V. harveyi were demonstrated. This result indicates that OmpW is a virulence related factor and it can be a vaccine candidate to prevent a high mortality caused by V. harveyi infection in olive flounder, Paralichthys olivaceus.

키워드

참고문헌

  1. Altschul S. F.. Madden T. L.. Schaffer A. A.. Zhang J.. Zhang Z.. Miller W. & Lipman D. J.(1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25, 3389-3402. https://doi.org/10.1093/nar/25.17.3389
  2. Alves N.. Jr, Neto O.. S. Silva B.. S. De Moura R.. L. Francini-Filho R. B.. Barreira E.. Castro C.. Paranhos R.. Bitner-Mathe B. C.. Kruger R. H.. Vicente A. C.. Thompson C. C. & Thompson F. L.(2010). Diversity and pathogenic potential of vibrios isolated from Abrolhos Bank coral. Environ Microbiol Rep 2(1), 90-95. https://doi.org/10.1111/j.1758-2229.2009.00101.x
  3. Austin B. & Zhang X.(2006). Vibrio harveyi: a significant pathogen of marine vertebrates and in vertebrates Lett Appl Microbiol 43, 119-124. https://doi.org/10.1111/j.1472-765X.2006.01989.x
  4. Edward J. N.(1996). Fish diseases: Diagnosis and treatment. Iowa state university Press, USA
  5. Ellis T. N. & Kuehn M. J.(2010). Virulence and immunomodulatory roles of bacterial outer membrane vesicles. Microbiol Mol Biol Rev 74, 81-94. https://doi.org/10.1128/MMBR.00031-09
  6. Fukuda Y.. Matsuoka S.. Mizuno Y. & Narita K.(1996). Pasteurella piscicida infection in cultured juvenile Japanese flounder. Fish Pathol 31, 33-38. https://doi.org/10.3147/jsfp.31.33
  7. Gil F.. Ipinza F.. Fuentes J.. Fumeron R.. Villarreal J. M.. Aspee A.. Mora G. C.. Vasquez C. C. & Saavedra C.(2007). The OmpW(porin) gene mediates methyl viologen (paraquat) efflux in Salmonella enterica serovar typhimurium. Res Microbiol 158, 529-536. https://doi.org/10.1016/j.resmic.2007.05.004
  8. Goel A. K. & Jiang S. C.(2010). Genetic determinants of virulence, antibiogram and altered biotype among the Vibrio cholerae O1 isolates from different cholera outbreaks in India. Infect Genet Evol 10, 815-819.
  9. Guex N. & Peitsch M. C.(1997). SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 18, 2714-2723. https://doi.org/10.1002/elps.1150181505
  10. Hong H.. Patel D. R.. Tamm L. K. & Berg BVD(2006) The outer membrane protein OmpW forms an eight-stranded beta-barrel with a hydrophobic channel. J Biol Chem 281, 7568-7577. https://doi.org/10.1074/jbc.M512365200
  11. Kim D. H.. Han H. J.. Kim S. M.. Lee D. C. & Park S. I.(2004). Bacterial enteritis and the development of the larval digestive tract in olive flounder Paralichthys olivaceus.J Fish Dis 27, 497-505. https://doi.org/10.1111/j.1365-2761.2004.00553.x
  12. Kim J. W.. Jung S. H.. Park M. A.. Do J. W.. Choi D. L.. Jee B. Y.. Cho M. Y.. Kim M. S.. Choi H. S.. Kim Y. C.. Lee J. S.. Lee C. H.. Bang J. D..Park M. S. & Seo J. S.(2006). Monitoring of pathogens in cultured fish of Korea for the summer period from 2000 to 2006. J Kor Fish Pathol 19, 207-214.
  13. Kim M. S.. Cho J. Y. & Choi H. S.(2014a). Identification of Vibrio harveyi, Vibrio ichthyoenteri, and Photobacterium damselae isolated from olive flounder Paralichthys olivaceus in Korea by multiplex PCR developed usingthe rpoB gene. Fish Sci 80, 333-339. https://doi.org/10.1007/s12562-014-0702-5
  14. Kim M. S.. Jung S. H. & Hong S.(2014b). Effect of 2-2'-dipyridyl in culture media and combined advantage of Streptococcus parauberis vaccine for preparation of Vibrio harveyi vaccine on olive flounder, Paralichthys olivaceus. JFMSE 72, 1366-1372.
  15. Lavilla-Pitogo C. R.. Baticados M. C. L.. Cruz-Lacierda E. R. & Pena L. D.(1990). Occurrence of luminous bacterial disease of Penaeus monodon larvae in the Philippines. Aquaculture 91, 1-13. https://doi.org/10.1016/0044-8486(90)90173-K
  16. Letunic I, Doerks T & Bork P(2012). SMART7: recent updates to the protein domain annotation resoutce. Nucleic Acids Res 40(D1), D302-D305. https://doi.org/10.1093/nar/gkr931
  17. Li K.. Gu W.. Liang J.. Xiao Y.. Qiu H.. Yang H.. Wang X. & Jing H.(2014). Gene polymorphism analysis of Yersinia enterocolitica outer membrane protein A and putative outer membrane protein A family protein. BMC Genomics 15, 201-208. https://doi.org/10.1186/1471-2164-15-201
  18. Mao Z.. Yu L.. You Z.. Wei Y. & Liu Y.(2007). Cloning, expression and immunogenicty analysis of five outer membrane proteins of Vibrio parahaemolyticus zj2003. Fish Shellfish Immunol 23(3), 567-575. https://doi.org/10.1016/j.fsi.2007.01.004
  19. Marhual N. P.. Samal S. K. & Das B. K.(2009). Random amplified polymorphic DNAPCR typing of Vibrio parahaemolyticus isolated from black tiger shrimp Penaeus monodon. E-planet, 7, 1-6.
  20. Marhual, N. P. . Das B. K. . Pradhan J. . Swain P. Mishra B. K. & Amberkar E.(2012). RAPD-PCR and outer membrane protein characterization of Vibrio alginolyticus and V. parahaemolyticus isolated from diseased shrimp. Israeli J. Aquacult.- Bamidgeh, IJA_64, 683-694.
  21. Muroga K.. Yasunobu H.. Okada N. & Masumura K.(1990). Bacterial enteritis of cultured flounder Paralichthys olivaceus larvae. Dis Aquat Org 9, 121-125. https://doi.org/10.3354/dao009121
  22. Nandi B.. Nandy R. K.. Mukhopadhyay S.. Nair G. B.. Shimada T. & Ghose A. C.(2000). Rapid method for species-specific identification of Vibrio cholerae using primers targeted to the gene of outer membrane protein OmpW. J Clin Microbiol 38(11), 4145-4151.
  23. Ningqiu L.. Junjie B.. Shuqin W.. Xiaozhe F.. Haihua L.. Xing Y. & Cunbin S.(2008). An outer membrane protein, OmpK, is an effective vaccine candidate for Vibrio harveyi in orangespotted grouper(Epinephelus coioides). Fish and Shellfish Immunology, 25, 829-833. https://doi.org/10.1016/j.fsi.2008.09.007
  24. Qian R.. Chu W.. Mao Z.. Zhang C.. Wei Y. & Yu L.(2007). Expression, characterization and immunogenicity of a major outer membrane protein from Vibrio alginolyticus. Acta Biochim Biophys Sin(Shanghai) 39(3), 194-200.
  25. Qian R.. Xiao Z.. Zhang C.. Chu W.. Mao Z. & Yu L.(2008). Expression and purification of two major outer membrane proteins from Vibrio alginolyticus. World J Microbiol Biotechnol 24, 245-251. https://doi.org/10.1007/s11274-007-9463-y
  26. Remmert M.. Biegert A.. Hauser A. & Soding J.(2012). HHblits: lightning-fast iterative protein sequence searching by HMM-HMM alignment. Nat Methods 9, 173-175.
  27. Sunaryanto A. & Mariam A.(1986). Occurrence of pathogenic bacteria causing luminosense in penaeid larvae in Indonesian hatcheries. Bull Brackishwater Aquaculture 8, 105-112.
  28. Tamura K.. Peterson D.. Peterson N.. Stecher G.. Nei M. & Kumar S.(2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28, 2731-2739. https://doi.org/10.1093/molbev/msr121
  29. Won K. M.. Choi J. H. & Park S. I.(2006). Characteristics of the extracellular products(ECPs) of Vibrio harveyi grown under various conditions. J Kor Fish Pathol 19, 119-126.
  30. Xu C.. Wang S.. Ren H.. Lin X.. Wu L. & Peng X.(2005). Protein analysis on the expression of outer membrane proteins of Vibrio alginolyticus at different sodium concentrations. Proteomics 5, 3142-3152. https://doi.org/10.1002/pmic.200401128
  31. Yu L.. Hu Y.. Sun B. & Sun L.(2013). Immunological study of the outer membrane proteins of Vibrio harveyi: Insights that link immunoprotectivity to interference with bacterial infection. Fish Shellfish Immunol 35, 1293-1300. https://doi.org/10.1016/j.fsi.2013.07.043