| Project/Area Number |
21810022
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Risk sciences of radiation/Chemicals
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| Research Institution | Nagasaki University |
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Principal Investigator |
NAKAZAWA Yuka Nagasaki University, 医歯薬学総合研究科, COE研究員 (00533902)
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| Project Period (FY) |
2009 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥2,652,000 (Direct Cost: ¥2,040,000、Indirect Cost: ¥612,000)
Fiscal Year 2010: ¥1,261,000 (Direct Cost: ¥970,000、Indirect Cost: ¥291,000)
Fiscal Year 2009: ¥1,391,000 (Direct Cost: ¥1,070,000、Indirect Cost: ¥321,000)
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| Keywords | DNA修復 / ヌクレオチド除去修復 / 紫外線 / エチニルデオキシウリジン / エチニルウリジン / In Cell Analyzer |
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| Research Abstract |
Nucleotide excision repair (NER) removes the major UV-photolesions from cellular DNA. Two assays commonly used in measurement of NER activity are 'unscheduled DNA synthesis (UDS)', and 'recovery of RNA synthesis (RRS)'. Reliable methods for these assays involved measurements of incorporation of radio-labeled nucleosides. We have established non-radioactive procedures for determining UDS and RRS levels by incorporation of alkyne-conjugated nucleoside analogues, 5-ethynyl-2'-deoxyuridine (EdU) and 5-ethynyuridine (EU). We assessed the UDS and RRS levels of several fibroblasts derived from NER-deficient patients by this non-radioactive assay system. Our results were in good agreement with those obtained by conventional assays, indicating that the system provides a convenient, but sensitive and accurate assay. We also demonstrated the utility of this technique for a large-scale siRNA-screening to identify novel NER factors.
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