| Research Abstract |
Toxic cyanobacterial (blue-green algal) waterblooms are found worldwide in eutrophic lakes and dams, and strains of several cyanobacterial genera produce the cyclic peptide hepatotoxin microcystins. The microcystins produced by cyanobacteria have become a serious environmental problem. In this studying, we developed three methods for detection of microcystins : (1) ELISA method, (2) Insecticidal bioassay method, (3) Gene probe and PCR method.
(1) ELISA method
Monoclonal antibodies against the microcystin LR variant were prepared from cloned hybridoma cell lines and ELISA method was developed.
(2) Insecticidal bioassay method
insecticidal bioassay for housefly was developed to investigate the toxicity of microcystin. The filter paper treated with microcystin in methanol was placed on a ethylene cup, then five adult houseflies were placed in the cup. After 24 hr, LD_<50> values of microcystin LR was 447 x 10^<-3> mu g/cm^2.
(3) Gene probe and PCR method
Microcystin synthetase genes were cloned and sequenced (ab. 45 kb). To amplify the microcystin synthetase genes, specific PCR primers were designed. Furthermore, specific DNA probes for detection of microcystin synthetase genes were designed. By using these probes and primers, microcystin-producing cells could be detected. Furthermore, regulation of gene expression in microcystin-producing Microcystis was revealed.
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