| Project/Area Number |
21K19039
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 37:Biomolecular chemistry and related fields
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| Research Institution | Gunma University |
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Principal Investigator |
Kamiya Koki 群馬大学, 大学院理工学府, 助教 (70612315)
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| Project Period (FY) |
2021-07-09 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2021: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
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| Keywords | ナノポアタンパク質 / リン脂質膜 / バイオセンシング / タンパク質改変 / 無細胞タンパク質合成系 / 膜タンパク質 / 一分子検出 / DNA検出 / リポソーム / 酵素反応 |
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| Outline of Research at the Start |
遺伝子組換えナノポアタンパク質を作製し様々な生体分子を認識できるシステムを構築を目指す。そして、構築した機構を用いバイオセンサと細胞機能制御に応用展開する。
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| Outline of Final Research Achievements |
We created a modified OmpG that expands and truncates β-hairpins, allowing the generation of small or large nanopores compared to that of wild-type (WT) OmpG nanopores. We demonstrated the detection of various structures of DNA (branched DNA) depending on the nanopore size using OmpG WT or mutated OmpG nanopores. Insights into the changes in pore diameters will be crucial to form precise pore diameters for the detection of various types of single biomolecules and for sequencing DNA, peptides, and proteins. Next, OmpG and OmpA synthesized by a cell-free synthesis system were incorporated into nano-sized liposomes. The nanopore formation in the nano-sized liposomes was confirmed using a patch-clamp method.
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| Academic Significance and Societal Importance of the Research Achievements |
βストランド数を増減させることでポア内のイオンの通過には、ポア内の水と接する部分のアミノ酸残基の電荷やかさ高さが重要な役割を果たしていることが分かった。改変体OmpGなどによって、分岐の長さが異なるT字DNAの検出に成功した。しがたって、自由にポアサイズを設計できるようになると、様々な大きさの生体分子の検出が可能になる。
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