Development of femtoliter chamber for single-molecule analysis of biological reaction

• Project/Area Number: 18201025
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Nanomaterials/Nanobioscience
• Research Institution: Osaka University
• Principal Investigator: NOJI Hiroyuki Osaka University, 産業科学研究所, 教授 (00343111)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥52,130,000 (Direct Cost: ¥40,100,000、Indirect Cost: ¥12,030,000); Fiscal Year 2008: ¥10,140,000 (Direct Cost: ¥7,800,000、Indirect Cost: ¥2,340,000); Fiscal Year 2007: ¥20,280,000 (Direct Cost: ¥15,600,000、Indirect Cost: ¥4,680,000); Fiscal Year 2006: ¥21,710,000 (Direct Cost: ¥16,700,000、Indirect Cost: ¥5,010,000)
• Keywords: 超微小溶液チャンバー / 1分子計測 / 回転分子モーター / 1分子DNA解析 / 制限酵素

Research Abstract

申請者が独自に開発した超微小溶液チャンバーを用いた1分子DNA解析技術の確立を中心に研究を行った。その結果、1分子制限酵素反応、1分子DNA増幅反応、1分子F1モーター反応効率計測に成功した。また、これまでのチャンバーとは異なり、溶液の添加・回収が可能なドロプレット型のチャンバーの開発も行った。これによって、酵素反応開始や細胞培養も可能となった。

Research Products

• [Journal Article] Correlation between the conformational states of F1-ATPase as determined from its crystal structure and single-molecule rotation (2008)
  • Author(s): Okuno D, Fujisawa R, Iino R, Hirono-Hara Y, Imamura H, Noji H
  • Journal Title: Proc. Natl. Acad. Soc. U.S.A 105 Pages: 20722-20727
  • Peer Reviewed
• [Journal Article] Photo gel-sol/sol-geltransition and its patterning of a supramolecular hydrogel as stimuli-responsive biomaterials (2008)
  • Author(s): Matsumoto S, Yamaguchi S, Ueno S, Komatsu H, Ikeda M, Ishizuka K, Iko Y, Tabata KV, Aoki H, Ito S, Noji H, Hamachi I
  • Journal Title: Chemistry 14(13) Pages: 3977-3986
  • Peer Reviewed
• [Journal Article] Loop-mediated isothermal amplification of a single DNA molecule in polyacrylamide gel-based microchamber (2008)
  • Author(s): Lam L, Sakakihara S, Ishizuka K, Takeuchi S, Arata HF, Fujita H, Noji H
  • Journal Title: Biomed Microdevices 10(4) Pages: 539-546
  • Peer Reviewed
• [Journal Article] Temperature-sensitivereaction intermediate of F1-ATPase (2008)
  • Author(s): Watanabe R, Iino R, Shimabukuro K, Yoshida M, Noji H
  • Journal Title: EMBO Rep 9(1) Pages: 84-90
  • Peer Reviewed
• [Journal Article] Lipid bilayer microarray for parallel recording of transmembrane ion currents (2008)
  • Author(s): Le Pioufle B, Suzuki H, Tabata KV, Noji H, Takeuchi S
  • Journal Title: Anal Chem 7(12) Pages: 1738-1745
  • Peer Reviewed
• [Journal Article] Correlation between the conformational states of F1-ATPase as determined from its crystal structure and single-mnolecule rotation (2008)
  • Author(s): Okuno D, Fujisawa R, Iino R, Hirono-Hara Y, Imamura H, Noji H
  • Journal Title: Proc. Natl. Acad. Soc. U. S. A 105 Pages: 20722-20727
  • Peer Reviewed
• [Journal Article] Photo gel-sol/sol-gel transition and its patterning of a supramolecular hydrogel as stimuli-responsive biomaterials (2008)
  • Author(s): Matsumoto S, Yamaguchi S, Ueno S, Komatsu H, Ikeda M, Ishizuka K, Iko Y, Tabata KV, Aoki H, Ito S, Noji H, Hamachi I.
  • Journal Title: Chemistry 14(13) Pages: 3977-3986
  • Peer Reviewed
• [Journal Article] Loop-nediated isothermal amplification of a single DNA molecule in polyacrylamide gel-based microchamber (2008)
  • Author(s): Lam L, Sakakihara S, Ishizuka K, Takeuchi S, Arata HF, Fujita H, Noji H
  • Journal Title: Biomed Microdevices 10(4) Pages: 539-546
  • Peer Reviewed
• [Journal Article] Temperature-sensitive reaction intermediate of F1-ATPase (2008)
  • Author(s): Watanabe R, Iino R, Shimabukuro K, Yoshida M, Noji H.
  • Journal Title: EMBO Rep. 9(1) Pages: 84-90
  • Peer Reviewed
• [Journal Article] Temperature-sensitive reaction intermediate of F(1)-ATP ase. (2008)
  • Author(s): Watanabe R, Iino R, Shimabukuro K, Yoshida M, Noji H.
  • Journal Title: EMBO Rep 9(1) Pages: 84-90
  • Peer Reviewed
• [Journal Article] Lipid bilayer microarray for parallel recording of transmembrane ion currents. (2008)
  • Author(s): Le Pioufle B, Suzuki H, Tabata KV, Noji H, Takeuchi S.
  • Journal Title: Anal Chem 80(1) Pages: 328-332
  • Peer Reviewed
• [Journal Article] An integrated system for enzymatic cleavage and electrostretching of freely-suspended single DNA molecules (2007)
  • Author(s): Lam L, Sakakihara S, Ishizuka K, Takeuchi S, Noji H
  • Journal Title: Lab Chip 7(12) Pages: 1738-1745
  • Peer Reviewed
• [Journal Article] Coupling of rotation and catalysis in F(1)-ATPase revealed by single-molecule imaging and manipulation (2007)
  • Author(s): Adachi K, Oiwa K, Nishizaka T, Furuike S, Noji H, Itoh H, Yoshida M,Kinosita K Jr
  • Journal Title: Cell 130(2) Pages: 309-321
  • Peer Reviewed
• [Journal Article] An integrated system for enzymatic cleavage and electrostretching of freely-suspended single DNA molecules. (2007)
  • Author(s): Lam L, Sakakihara S, Ishizuka K, Takeuchi S, Noii H.
  • Journal Title: Lab Chip 7(12) Pages: 1738-1745
  • Peer Reviewed
• [Journal Article] Coupling of rotation and catalysis in F(1)-ATPase revealed by single-molecule imaging and manipulation. (2007)
  • Author(s): Adachi K, Oiwa K, Nishizaka T, Furuike S, Noji H, Itoh H, Yoshida M, Kinosita K Jr.
  • Journal Title: Cell 130(2) Pages: 309-321
  • Peer Reviewed
• [Journal Article] Temperature distribution measurement on microfabricated thermodevice for singlebiomolecular observation using fluorescent dye (2006)
  • Author(s): Arata, H.F., Low, P., Ishizuka, K., Bergaud, C., Kim, B., Noji, H. and Fujita, H
  • Journal Title: Sensors and Actuators, B : Chemical 117(2) Pages: 339-345
  • Peer Reviewed
• [Journal Article] Chemical delivery microsystem for single-molecule analysis using multilaminar continuous flow (2006)
  • Author(s): Lee, S.W., Yamamoto, T.,Noji, H. and Fujii, T.
  • Journal Title: EBzyme and Microbial Technology 39(3) Pages: 519-525
  • Peer Reviewed
• [Journal Article] Motion control of single F1-ATPase rotary biomolecular motor using microfabricated local heating devices (2006)
  • Author(s): Arata, H.F.,Noji, H. and Fujita, H.
  • Journal Title: APPLIED PHYSICS LETTERS 88 Pages: 83902-83902
  • Peer Reviewed
• [Journal Article] Temperature distribution measurement on microfabricated thermodevice for single biomolecular observation using fluorescent dye. (2006)
  • Author(s): Arata, H.F., Low, P., Ishizuka, K., Bergaud, C., Kim, B., Noji, H., Fujita, H.
  • Journal Title: Sensors and Actuators, B : Chemical 117(2) Pages: 339-345
• [Journal Article] Chemical delivery microsystem for single-molecule analysis using multilaminar continuous flow (2006)
  • Author(s): Lee, S.W., Yamamoto, T., Noji, H., Fujii, T.
  • Journal Title: EBzyme and Microbial Technology 39 (3) Pages: 519-525
• [Journal Article] Motion control of single F1-ATPase rotary biomolecular motor using microfabricated local heating devices (2006)
  • Author(s): Arata, H.F., Noji, H., Fujita, H.
  • Journal Title: APPLIED PHYSICS LETTERS 88 Pages: 3-3
• [Presentation] Single molecule studies on F1-ATPase molecular motor (2009)
  • Organizer: 第16回International Colloquium on Scanning Probe Microscopy
  • Place of Presentation: Izu,atagawa(Japan)
  • Year and Date: 2009-12-13
• [Presentation] Single-molecule Imaging of ATP-driven Stepping Rotation of FoF1-ATP Synthase Reconstituted into Supported Membrane (2009)
  • Author(s): 飯野亮太 Khek-Chian Tham, 田端和仁 上野博史 野地博行
  • Organizer: Biophysical Society 53rd Annual Meting
  • Place of Presentation: アメリカ(ボストン)
  • Year and Date: 2009-03-01
• [Presentation] How is the Temperature Sensitive (TS)Reaction of F_1-ATPase Coupled with its Rotation? (2009)
  • Author(s): 榎佐和子 飯野亮太 渡辺力也 野地博行
  • Organizer: Biophysical Society 53rd Annual Meting
  • Place of Presentation: アメリカ(ボストン)
  • Year and Date: 2009-03-01
• [Presentation] Single molecule biophsicis of F1-ATPase : Mechanical mondulation od reaction rate constants and equilibrium constants by F1-ATPase (2009)
  • Organizer: 第6回Asian biophysics Association Symposium
  • Place of Presentation: HKUST(Hong Kong)
  • Year and Date: 2009-01-11
• [Presentation] Single molecule biophsicis of F1-ATPase : Mechanical mondulation od reaction rate constants and equilibrium constants by F1-ATPase (2009)
  • Author(s): 野地 博行
  • Organizer: 第6回Asian biophysics Association Symposium
  • Place of Presentation: HKUST(香港)
  • Year and Date: 2009-01-11
• [Presentation] Femtoliter chamber for Single-molecule and single-cell analysis (2009)
  • Organizer: 第4回IEEE-NEMS 09
  • Place of Presentation: Shinsen(China)
  • Year and Date: 2009-01-06
• [Presentation] Femtoliter chamber for Single-molecule and single-cell analysis (2009)
  • Author(s): 野地 博行
  • Organizer: 第4回IEEE-NEMS09
  • Place of Presentation: 中国(シンセン)
  • Year and Date: 2009-01-06
• [Presentation] Single molecule studies on Fl-ATPase molecular motor (2008)
  • Author(s): 野地 博行
  • Organizer: 第16回 International Colloquium on Scanning Pobe Microscopy
  • Place of Presentation: 伊豆・熱川
  • Year and Date: 2008-12-13
• [Presentation] 生命システムの階屑間をまたぐイメージング技術 (2008)
  • Author(s): 野地 博行
  • Organizer: BMB 2 0 0 8
  • Place of Presentation: 神戸国際展示場2号館3階
  • Year and Date: 2008-12-12
• [Presentation] 膜タンパク質ダイナミクス計測のための膜システムの開発 (2008)
  • Author(s): 野地 博行
  • Organizer: 第46回生物物理学会
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2008-12-04
• [Presentation] 超高感度バイオセンシング技術 (2008)
  • Author(s): 飯野 亮太
  • Organizer: 産業科学研究協会 平成20年度第3回産研テクノサロン
  • Place of Presentation: 大阪
  • Year and Date: 2008-10-30
• [Presentation] Mechanical modulation of ATP-binding and hydrolysis by single F1-ATPase molecule (2008)
  • Organizer: MPSA2008
  • Place of Presentation: Sapporo(Japan)
  • Year and Date: 2008-08-29
• [Presentation] Mechanical modulation of ATP-binding and hydrolysis by single F1-ATPase molecule (2008)
  • Author(s): 野地 博行
  • Organizer: MPSA2008
  • Place of Presentation: 札幌
  • Year and Date: 2008-08-29
• [Presentation] Mechanochemical coupling of F_1-ATPase and intracellular ATP imaging (2008)
  • Author(s): 飯野亮太 渡部力也 奥野大地 今村博臣 野地博行
  • Organizer: 15th European Bioenergetics Conference (EBEC2008)
  • Place of Presentation: イギリス(ダブリン)
  • Year and Date: 2008-07-19
• [Presentation] Single Molecule Studies on F1-ATPase (2008)
  • Organizer: International Symposium on Hierarchy and Holism "Bridging across Different Hierarchies in Natural Sciences"
  • Place of Presentation: Okazaki(Japan)
  • Year and Date: 2008-03-23
• [Presentation] Single Molecule Studies on F1-ATPase (2008)
  • Author(s): 野地 博行
  • Organizer: International Symposium on Hierarchy and Holism (ISHH), "Bridging across Different Hierarchies in Natural Sciences"
  • Place of Presentation: Okazaki (Japan)
  • Year and Date: 2008-02-23
• [Presentation] ATP-driven Rotation of FoF1-ATP Synthase Reconstituted into Supported Membrane (2008)
  • Author(s): 飯野 亮太
  • Organizer: 第52回米国生物物理学会年会
  • Place of Presentation: California (USA)
  • Year and Date: 2008-02-05
• [Presentation] Single-molcule Imaging of the transport-vesicle formation mediated by COPII system reconstituted on a planner bilayer system (2008)
  • Organizer: 第17回CDB会議(Organizer)
  • Place of Presentation: Kobe(Japan)
• [Presentation] Single-molcule Imaging of the transport-vesicle formation mediated by COPII system reconstituted on a planner bilayer system (2008)
  • Author(s): 野地 博行
  • Organizer: 第17回 CDB会議(野地オーガナイザー)
  • Place of Presentation: 理科学研究所(神戸)
• [Presentation] Single Molecule Studies on F1-ATPase (2007)
  • Author(s): 野地 博行
  • Organizer: The 6th JSPS Forum in France, "Chemical and physical nanobiology for medicine"
  • Place of Presentation: Strasbourg (France)
  • Year and Date: 2007-11-23
• [Presentation] Single Molecule Studies on F1-ATPase (2007)
  • Author(s): 野地 博行
  • Organizer: Autumn School for PhD students and young researcher (NAMIS) 2007
  • Place of Presentation: Tokyo (Japan)
  • Year and Date: 2007-11-05
• [Presentation] Single Molecule Studies on F1-ATPase (2007)
  • Author(s): 野地 博行
  • Organizer: Asian and Pacific Biomechanics (AP Biomech) 2007
  • Place of Presentation: Tokyo (Japan)
  • Year and Date: 2007-11-05
• [Presentation] Rapid detection of drug efflux from single bacterial cell enclosed in fem toliter chamber array (2007)
  • Author(s): 飯野 亮太
  • Organizer: μ TAS 2007
  • Place of Presentation: Paris (France)
  • Year and Date: 2007-10-09
• [Presentation] Versatile Acrylamide-based Microchambers for Single Molecular Biological Assays and Analysis (2007)
  • Author(s): Liza Lam
  • Organizer: μ TAS 2007
  • Place of Presentation: Paris (France)
  • Year and Date: 2007-10-09
• [Presentation] Single Molecule Studies on F1-ATPase (2007)
  • Author(s): 野地 博行
  • Organizer: 3rd work shop of the UK-Japan Bionanotechnology Collaboration
  • Place of Presentation: Oxford (UK)
  • Year and Date: 2007-07-19
• [Book] 細胞の回転エネルギー変換装置. 「超分子サイエンス」 (2009)
  • Author(s): 野地博行, 田端和仁, 飯野亮太
  • Publisher: NTS、編集中(in press)
• [Book] 第4章第6節. 培風館、編集中 (2009)
  • Author(s): 飯野亮太 野地博行
  • Publisher: 分子機械. 「基礎ナノバイオテクノロジー」(in press)
• [Book] 「超微小溶液チャンバーを利用した1分子バイオアッセイ」 (2008)
  • Author(s): 野地 博行
  • Publisher: 丸善
• [Book] 「分子マシンに新たな扉が開かれた」最新分子マシン. ナノで働く"高度な機械"を目指して (2008)
  • Author(s): 金原数、竹内昌治、竹内正之、野地博行
  • Publisher: 化学同人
• [Book] 超微笑溶液チャンバーを利用した1分子バイオアッセイ (2008)
  • Author(s): 野地 博行
  • Total Pages: 6
  • Publisher: 丸善
• [Book] 分子マシンに新たな扉が開かれた (2008)
  • Author(s): 金原数, 竹内昌治, 竹内正之, 野地博行
  • Total Pages: 11
  • Publisher: 化学同人
• [Book] F1-ATPaseに見る回転分子モーターのナノバイオロジー (2008)
  • Author(s): 野地博行, 田端和仁
  • Total Pages: 10
  • Publisher: 化学同人
• [Book] 最新分子マシン. ナノで働く“高度な機械"を目指して (2008)
  • Author(s): 飯野 亮太
  • Total Pages: 2
  • Publisher: 化学同人
• [Book] 未来材料「超微小反応チャンバーを用いた高感度バイオアッセイ」 (2007)
  • Author(s): 飯野 亮太, Liza Lam, 野地 博行
  • Total Pages: 68
  • Publisher: 未来材料
• [Book] 生物物理学ハンドブック (2007)
  • Author(s): 野地 博行
  • Total Pages: 680
  • Publisher: 朝倉書店
• [Book] ナノバイオ計測の実際 (2007)
  • Author(s): 野地 博行
  • Total Pages: 206
  • Publisher: 講談社
• [Book] ナノバイオ大辞典 (2007)
  • Author(s): 野地 博行
  • Total Pages: 636
  • Publisher: テクノシステム
• [Book] 蛋白質 核酸 酵素Vol.52 No.4 (2007)
  • Author(s): 野地 博行
  • Total Pages: 396
  • Publisher: 共立出版
• [Book] ナノテクのためのバイオ入門 (2007)
  • Author(s): 野地 博行
  • Total Pages: 224
  • Publisher: 共立出版
• [Book] 細胞工学 (2006)
  • Author(s): 野地 博行
  • Total Pages: 134
  • Publisher: 秀潤社
• [Book] 生物物理 (2006)
  • Author(s): 野地 博行
  • Total Pages: 55
  • Publisher: 日本生物物理学会
• [Book] Micro Total Analysis Systems 2006 (2006)
  • Author(s): Lam, L., Ishizuka, K., Sakakihara, S., Noji, H.
  • Total Pages: 560
  • Publisher: 化学とマイクロ・ナノシステム研究会
• [Patent(Industrial Property Rights)] 細胞検体の異物排出活性検出方法、及びその利用 (2006)
  • Inventor(s): 飯野亮太, 西野邦彦, 仲田昌義, 榊原昇一, 山口明人, 野地博行
  • Industrial Property Rights Holder: 国立大学法人大阪大学
  • Industrial Property Number: 2006-294558
  • Filing Date: 2006-10-30