1997 Fiscal Year Final Research Report Summary
Immunological methods of identification of microorganisms producing cancer promotors in fresh water and a detection method of the promotor
| Project/Area Number |
07660276
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | Fukui Prefectural University |
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Principal Investigator |
HIROISHI Shingo Fukui Prefectural University, Faculty of Biotechnology, Professor, 生物資源学部, 教授 (00114190)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Microcystis / monoclonal antibody / identification |
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| Research Abstract |
Many microorganisms were reported to cause blooms in several aquatic sreas in the world. Microcystis is one of the most important microorganisms among them producing microcystins which has hepato-toxicity and hepato-carcinogenic promoter activity.
Species belonging to the genus are mainly identified by morphology such as shape of colony. However, their classification by morphology is quite difficult when colony formation of these microorganisms are insufficient dependent on environmental conditions. Therefore, we tried to identify them by recognizing antigen molecules distributed on the surface of the cells using monoclonal antibodies.
Female BALB/c mice (4 weeks of age) were immunized with Microcystis cell lines. The spleen cells of the mice were fused with cultured myeloma cells by polyethylene glycol. Reactivities of the monoclonal antibodies produced by the hybridomas obtained by cell fusion were detected by an indirect fluorescence technique.
We developed 3 hybridomas producing antibodies and examined their specificity of reactivity using 38 strains of Microcystis. As a result, they were specifically reactive with groups of Microcystis classified by 16S rDNA analysis rather than morphology. Our method might be more reliable than that by morphology.
One of the monoclonal antibodies obtained was reactive with the cells fixed with glutaraldehyde, and with those at both exponentially and stationary phases.
Flowcytemetric analysis using those antibodies revealed that this analysis could be useful for quantitative analysis of field samples.
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