Abstract
Vibrio harveyi, the major causative agent of vibriosis, affects a diverse range of marine cultured organisms over a wide geographical area. However, reports about screening the effective antigen and research on vaccines of V. harveyi are scarce. Flagellin, lipopolysaccharide (LPS) and outer membrane proteins (OMP) are major immunogenic antigens in many Gram-negative bacteria. In this study, the flagellin, OMP and LPS of the V. harveyi TS-628 strain isolated from infected groupers were extracted and Western blot analysis was used to detect the antigenicity of these extractions. Results of the Western blot assay reveal that there are four positive flagellin bands: 35 kDa, 38 kDa, 43 kDa, and 52 kDa, of which the 43 kDa and 52 kDa bands displayed the strongest positive reaction. There are five positive OMP bands about 35 kDa, 38 kDa, 43 kDa, 47 kDa, and 52 kDa, of which the 43 kDa appeared to have the strongest positive reaction although the other four proteins also displayed strong reactions. However, LPS is Western blot-negative. These results indicate that the 43 kDa and 52 kDa flagellin and OMP of size 43 kDa, 52 kDa can be candidates for developing vaccines against V. harveyi.
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References
Chen C F, Chen C R (2002). Immunogenicity of crude lipopolysaccharide of three pathogenic bacteria of fish to silver crucian carp, Caressius Auratus Gibelio. Acta Hydrobiologica Sinica, 26(5): 483–488 (in Chinese)
Chen X G, Wu S Q, Shi C B, Li N Q (2004). Isolation and identification of pathogenetic Vibrio harveyi from estuary cod Epinephelus coioides. Journal of Fishery Sciences of China, 2004, 11(4): 313–317 (in Chinese)
Das M, Chopra A K, Wood T, Peterson J W (1998). Cloning, sequencing and expression of the flagellin core protein and other genes encoding structural proteins of the Vibrio cholerae flagellum. FEMS Microbiology Letters, 165: 239–246
Gou X J (2001). Electrophoresis An Experimental Technology for Protein. Beijing: Science Press (in Chinese)
Lin K B, Zhou C, Liu J F (1999). Studies on the pathogenic bacteria of Pseudosciaena crocea in marine cage culture. Marine Sciences, 4: 58–62 (in Chinese)
Liu P C, Lee K K, Tu C C, Chen S N (1997). Purification and characterization of a cysteine protease produced by pathogenic luminous Vibrio harveyi. Curr Microbiol, 35: 32–39
Lowery C A, McKenzie K M, Qi L, Meijler M M, Janda K D (2005). Quorum sensing in Vibrio harveyi: Probing the specificity of the LuxP binding site. Bioorganic and Medicinal Chemistry Letters, 15: 2395–2398
McCarter L L (1995). Genetic and molecular characterization of the polar flagellum of Vibrio parahaemolyticus. Journal of Bacteriology, 77(6): 1595–1609
McDonough M W, Smith S E (1976). Molecular weight variation among bacterial flagellins. Microbiology, 16(63): 49–53
McGee D, Horstedt P, Milton D L (1996). Identification and characterization of additional flagellin genes from Vibrio anguillarum. Journal of Bacteriology, 178(17): 5188–5198
Milton D L, O’Toole R, Hörstedt P, Wolf-Watz H (1996). Flagellin A is essential for the virulence of Vibrio anguillarum. Journal of Bacteriology, 1310–1319
Ming T C, Frasch C E (1982). A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Analytical Biochemistry, 119(1): 119–115
Nikaido H (1997). Isolation of outer membranes. In: Patrik L C, ed. Bacterial Pathogenesis. London: Academic Press, 113–122
Qin Y X, Wang J, Su Y Q, Wang D X and Chen X Z (2006). Studies on the Pathogenic Bacterium of Ulcer Disease in Epinephelus awoara. Acta Oceanologica Sinica, 25(1): 147–153
Saeed M O (1995). Association of Vibrio harveyi with mortalities in cultured marine fish in Kuwait. Aquaculture, 136(1): 21–29
Sambrook J, Russel D W (2002). Molecular Cloning: A Laboratory Manual. 3rd ed. Beijing: Science Press
Shen P (2000). Microbiology. Beijing: Higher Education Press (in Chinese)
Su B J, Nie P (2001). Outer membrane protein and lipopolysaccharide of Cytophaga columnaris and their immunogenicity to the mandarin fish, Siniperca chuatsi. Acta Hydrobiologica Sinica, 25(5): 524–527 (in Chinese)
Suzuki S, Kuroe K, Yasue K (1996). Antigenicity and N-terminal amino acid sequence of a 35kD porin-like protein of Listonella(Vibrio) anguillarum: Comparison among different serotypes and other bacterial species. Lett Appl Microbio, 23: 257–260
Tendencia E A, dela Pena M R, Fermin A C, Lio-Po G, Choresca Jr C H, Inui Y (2004). Antibacterial activity of tilapia Tilapia hornorum against Vibrio harveyi. Aquaculture, 232: 145–152
Totten P A, Lory S L (1990). Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK. Journal of Bacteriology, 172(12): 7188–7199
Tu X L, Kawai K (1999). Antigenic profile and protective role of a 37kDa major outer membrane protein of Edwardsiella tarda. Fish Pathol, 34(2): 59–64
Ulrich D L, Kojetin D, Bassler B L, Cavanagh J, Loria J P (2005). Solution structure and dynamics of LuxU from Vibrio harveyi, a phosphotransferase protein involved in bacterial quorum sensing. Journal of Molecular Biology, 347: 297–307
Wang G L, Jin S (2000). Studies on the skin ulcer disease and pathogenic bacterium of marine cage-cultured sea-perch. Journal of Oceanography of Huanghai and Bohai Seas, 18(3): 85–89 (in Chinese)
Westphal O, Jann K (1965). Bacterial lipopolysaccharides: Extraction with phenol-water and further applications of the procedure. Methods in Carbohydrate Chemistry, 5: 83–96
Wu H B, Pan J P (2001). Progress in studies of vibriosis in aquaculture. Journal of Fishery Sciences of China, 8(1): 89–93 (in Chinese)
Yang G C, Schrank G D, Freeman B A (1977). Purification of flagellar cores of Vibrio cholerae. Journal of Bacteriology, 192(2): 1121–1128
Yu Y R, Liu X C (2000). Progress of studies on membrane protein of Gram-negative bacteria. Progress in Veterinary Medicine, 21(2): 35–39 (in Chinese)
Zhang X H, Rbertson P, Austin B, Xu H (1997). Comparison of outer membrane protein profiles of Vibrio sp.. Acta Microbiologica Sinica, 37(6): 449–454 (in Chinese)
Zhang X H, Xu H X (1997). Studies on antigenicity of outer membrane proteins of Vibrio parahemolyticus. Journal of Fishery Sciences of China, 4(4): 49–53 (in Chinese)
Zhou L, Liu H M, Zhan W B (2003). Isolation and characteristics of major outer membrane proteins of aquatic pathogens Vibrio anguillarum and Vibrio alginolyticus. Journal of Fishery Sciences of China, 10(1): 31–35 (in Chinese)
Zhu L P, Chen X Q (2002). The Usual Experimental Methods of Immunology. Beijing: People’s Surgeon Publishers (in Chinese)
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Translated from Journal of Xiamen University (Natural Science), 2006, 45(3): 393–396 [译自: 厦门大学学报 (自然科学版)]
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Qin, Y., Wang, J., Wang, S. et al. Antigenicity analysis of Vibrio harveyi TS-628 strain. Front. Biol. China 2, 263–267 (2007). https://doi.org/10.1007/s11515-007-0037-z
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DOI: https://doi.org/10.1007/s11515-007-0037-z