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http://hdl.handle.net/10119/14334
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Title: | 光架橋反応を用いた19FーNMRによる核酸類検出法の開発 |
Other Titles: | Development about photochemical detection of nuceic acids by 19F-NMR |
Authors: | 藤本, 健造 |
Authors(alternative): | Fujimoto, Kenzo |
Keywords: | 19FーNMR 核酸検出 ケミカルシフト |
Issue Date: | 1-Jun-2017 |
Abstract: | 19F NMRのケミカルシフトを用いた核酸類検出法は、生体バックグラウンドが皆無であり、高い生体透過性を有することから、生体内などの夾雑試料中での核酸検出法として注目されている。本研究では、夾雑試料中でもB/F分離可能な大きなケミカルシフトの変化(8.0 ppm)を誘起可能な新規核酸類検出法(19F NMRを用いた核酸類検出法)を開発した。さらに、実際の生体内に存在する核酸濃度は低い為、増幅系を組み込んだ配列選択的な核酸類検出法を開発し、最終的にnmスケールの核酸類の検出にも成功した。:We demonstrated an approach for the chemical shift-based detection of nucleic acids based on CNVK (cyanovinylcarobazole) mediated DNA/RNA photo-cross-linking. The 19F NMR signal of TFT(trifluoromethyl thymidine) in a DNA duplex structure was dramatically shifted from -63.2 ppm to -71.2 ppm induced by photo-cross-linking. This larger chemical shift change induced by CNVK mediated DNA photo-cross-linking enables DNA and RNA detection without confounding effects due to the influence of neighboring bases. We have also successfully demonstrated that 10 nM miRNA can be detected in a sequence-specific manner based on miRNA amplification and chemical shift caused by the photo-cross-linking of CNVK and TFT in DNA and RNA. |
Description: | 基盤研究(B)(一般) 研究期間:2014~2016 課題番号:26288077 研究者番号:90293894 研究分野:核酸化学 |
Language: | jpn |
URI: | http://hdl.handle.net/10119/14334 |
Appears in Collections: | 2016年度 (FY 2016)
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26288077seika.pdf | | 455Kb | Adobe PDF | View/Open |
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