| Budget Amount *help |
¥11,600,000 (Direct Cost: ¥11,600,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2001: ¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 2000: ¥4,200,000 (Direct Cost: ¥4,200,000)
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| Research Abstract |
Due to the high demand for the standard material and new scientific tools in the field of research related to paralytic shellfish toxins, we developed the new nethods and prepared several saxitoxin (STX) derivatives by chemical conversions. Mass culture of toxigenic cyanobacteria Anabaena circinalis was used to obtained starting materials, C1, C2, gonyautoxin-2 and 3 (GTX2,3). As HPLC standards, dcGYX2, dcGYX3, dcSTX and GTX5, which were difficult to obtain from naturally contaminated shellfish, were derivatized by simple methods from C1, C2. DcSTX was shown to work as an alternate for STX as standard for the mouse bioassay. Highly effective method to prepare acetyldecarbamoylsaxiton was developed as a model for radio-labeled STX. Two types of affinity gels were also prepared to utilize for the research on macromolecule involved in the accumulation and transportation of toxins in bivalves and for the purification of voltage gated sodium channel protein. First the moieties having carboxyl were introduced at C11 and C13 of STX skeleton, namely 11-(2-carboxyethylthio) saxitoxin and 13-O-hemisuccinyl decarbamoylsaxitoxin. They were coupled with agarose gel (Sepharose 4B) via spacer. The former affinity gel was proven to be effective in purification of STX-binding protein in puffer plasma and to be stable in storage at 4℃ for more than 6 months.
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