| Project/Area Number |
14360111
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General fisheries
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| Research Institution | Kochi University |
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Principal Investigator |
KAWAI Kenji Kochi University, Faculty of Agriculture, Professor, 農学部, 教授 (60127925)
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| Co-Investigator(Kenkyū-buntansha) |
OSHIMA Syunichirou Kochi Univ., Graduate School of Kuroshio Science, Associate Professor, 黒潮圏海洋科学研究科, 助教授 (80325406)
HOSOKAWA Hidetsuyo Kochi University, Faculty of Agriculture, Professor, 農学部, 教授 (40036744)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2004: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2003: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2002: ¥7,400,000 (Direct Cost: ¥7,400,000)
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| Keywords | edwardsiellosis / outer membrane protein / common antigen / vaccine / fish disease / ricombinant / GAPDH / Edwardsiella tarda / 血清型 |
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| Research Abstract |
As there are many serotypes in fish pathogenic bacterium Edwardsiella tarda it has been considered difficult to produce effective vaccine against this disease. Present research project is intended to develop an.vaccine based on a commonly protective antigen among the different serotypes of E.tarda and obtained following results.
A 37kDa outer membrane protein(OMP) was found as a common antigen among different serotypes of E.tarda. This protein was detected as separated one band by general SDS-PAGE and Western blotting. Two-dimensional electrophoresis and Western blotting showed the 37kDa OMP was detected as separate several dots with different PIs. However, these proteins showed similar antigenicity and same amino acid sequences, which indicate small difference in the molecular structure due to phosphonium or glycosyl modifications. N-terminal amino acid sequence of the protein and complete nucleotide sequence of the gene demonstrated that the 37kDa OMP is a glyceraldehyde-3-phosphate dehydrogenase (GAPDH) found in a variety of bacteria. The recombinant GAPDH (rGAPDH) of E. tarda over expressed by using a Escherichia coli expression system was immunized to Japanese flounder and evaluated its vaccine efficacy. The results showed that the immunization with rGAPDH effectively protected Japanese flounder against infection with the different serotypic strains of E.tarda. The immune serum of these fish demonstrated increase in the antibody titer against those strains. The results show that the use of the rGAPDH will enable to develop a commonly effective vaccine against any serotypes of E.tarda.
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