| Project/Area Number |
15380144
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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 |
Fisheries chemistry
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| Research Institution | Kitasato University |
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Principal Investigator |
KODAMA Masaaki Kitasato University, School of Fisheries Sciences, Professor, 水産学部, 教授 (40050588)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Shigeru Kitasato University, School of Fisheries Sciences, Associate Professor, 水産学部, 助教授 (20170748)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥15,700,000 (Direct Cost: ¥15,700,000)
Fiscal Year 2005: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2004: ¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 2003: ¥9,400,000 (Direct Cost: ¥9,400,000)
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| Keywords | PSP / saxitoxin / endocellular bacteria / saxitoxin-producing bacteria / anti-szxitoxin antibody / conjugate of saxitoxins and thiols / saxitoxin-protein cimplex / non-specific protease / 抗麻ひ性貝毒抗体 / ELISA / 麻ひ性貝毒結合タンパク / 麻ひ性貝毒生産細菌 / Alexandrium tamarense / Moraxella / western blotting / protease / 細菌 / 抗体 / 中間代謝物 / 毒結合タンパク |
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| Research Abstract |
PSP toxins are toxins produced by toxic dinoflagellates such as Alexandrium tamarense. On the contrary, we suggested that toxins originate from endocellular bacteria of dinoflagellates, based on the facts that endocellular bacteria of A. tamarense produce saxitoxins, though the productivity is small. We propose here the hypothesis that bacteria produce precursor of toxins which is changed to toxins in dinoflagellates, Based on the hypothesis, we tried to find the precursor substance in bacterial. First of all, the specific antibody against saxitoxins which detects substance(s) possessing ligand with saxitoxin structure, by development of specific antibody against saxitoxin with the antigen utilizing the reaction between toxins and thiols. Western blot analysis of bacterial proteins by the antibody showed the protein bands stained by the antibody, indicating that these proteins possess ligands with saxitoxin structure. Then, the bacterial protein fraction was digested with nonspecific protease to obtain the digested protein fragments. ELISA using the antibody showed that low molecular fragment in the hydrolyzate reacts with the antibody. Trial to isolate the ELISA-positive substance showed the difficulty to isolate the substance, because of contamination of many impurities contaminated in the hydrolyzed. On the other hand, the substance was found to be effectively isolated, just in one step, when immunoprecipitation using the antibody was applied. Isolation of the substance for structure identification is on the way.
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