| Project/Area Number |
15K08612
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | University of Toyama |
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Principal Investigator |
Kitajima Isao 富山大学, 大学院医学薬学研究部(医学), 教授 (50214797)
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| Co-Investigator(Kenkyū-buntansha) |
仁井見 英樹 富山大学, 大学院医学薬学研究部(医学), 准教授 (50401865)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | 転写因子 / ＮＦ－κＢ / 蛍光相関分光法 / ＥＬＩＳＡ / 活性定量 / プロファイル解析 / ＡＰ１ / DNA-転写因子結合アレイ法 / 先端検査デバイス / 炎症 / 発現プロファイル / CAGE法 |
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| Outline of Final Research Achievements |
As for the quantity transcription factor activated analysis, an ELISA method is used at a study level. We developed new experimental apparatus for the detection of inflammatory allied transcription factor activity based on fluorescence correlation spectroscopy (FCS), which is able to analyze binding between DNA and protein in the liquid phase. We made each fluorescence DNA probe which specifically bound to NF-κB, AP1, NF-IL-6, SP1, NF-AT or OCT1. To assess the applicability of the FCS method, we quantitated each transcription factor activity in the human lymphocytes stimulated with TNF-α, LPS, or PMA/Ionomycin. In results, 1.5-fold increased NF-κB and AP1 activity was measured two hours after stimulation with TNF-α or PMA/Ionomycin. The quantity transcription assay by the FCS method is quick, simple, easy and high-throughput analytical method, compared to the ELISA method.
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