Project/Area Number |
11470498
|
Research Category |
Grant-in-Aid for Scientific Research (B).
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Environmental pharmacy
|
Research Institution | Kanazawa University |
Principal Investigator |
NIKAIDO Osamu Kanazawa University, Faculty of Pharmaceutical Sciences, Professor., 薬学部, 教授 (60019669)
|
Co-Investigator(Kenkyū-buntansha) |
MATSUNAGA Tsukasa Kanazawa University, Faculty of Pharmaceutical Sciences, Associate Professor., 薬学部, 助教授 (60192340)
石垣 靖人 金沢大学, 薬学部, 助手 (20232275)
|
Project Period (FY) |
1999 – 2000
|
Project Status |
Completed (Fiscal Year 2000)
|
Budget Amount *help |
¥15,400,000 (Direct Cost: ¥15,400,000)
Fiscal Year 2000: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥12,200,000 (Direct Cost: ¥12,200,000)
|
Keywords | DNA damage / cyclobutane pyrimidine dimer (CPD) / (6-4) photoproduct / Capillary electrophoresis / Enzyme-linked immunosorbent assay (ELISA) / Dewar photoproduct / monoclonal antibody / Dot-blot analysis / シクロブタン型ビリミジン二量体 / ヌクレオチド除去修復 / シクロブンタン型ピリミジンニ量体 / 蛍光標識抗体 |
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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