2000 Fiscal Year Final Research Report Summary
Establishment and application of the sensitive assay systems for ultraviolet-induced DNA damage.
| Project/Area Number |
11470498
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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 |
Environmental pharmacy
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| Research Institution | Kanazawa University |
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Principal Investigator |
NIKAIDO Osamu Kanazawa University, Faculty of Pharmaceutical Sciences, Professor., 薬学部, 教授 (60019669)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUNAGA Tsukasa Kanazawa University, Faculty of Pharmaceutical Sciences, Associate Professor., 薬学部, 助教授 (60192340)
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| Project Period (FY) |
1999 – 2000
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| Keywords | DNA damage / cyclobutane pyrimidine dimer (CPD) / (6-4) photoproduct / Capillary electrophoresis / Enzyme-linked immunosorbent assay (ELISA) / Dewar photoproduct / monoclonal antibody / Dot-blot analysis |
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| Research Abstract |
We established the quantitative systems to detect various kinds of ultraviolet (UV)-induced DNA damage by using,
1) a capillary electrophoresis combined with laser-induced fluorescence detector. The DNA irradiated with various doses of UVC was incubated with the first antibody recognizing DNA damage, followed by incubation with the second antibody (anti-mouse IgG goat antibody) conjugated with fluorescent dye. The DNA damage-first antibody-second antibody-complex was detected in a UVC dose-dependent manner in the rage of 0-1J/m^2.This higher resolution of the capillary electrophoresis for the detection of cyclobutane pyrimidine dimers (CPD) in UV-irradiated DNA made us possible to assess the biological effect of physiologically relevant doses of ultraviolet light in solar light.
2) an enzyme-linked immunosorbent assay (ELISA) conjugated with enhancedchemiluminescence system (ECL). We successfully detected CPD in DNA irradiated with UVC doses up to 1J/m^2 by this ECL-ELISA system.
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