• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development and application of Giga-screening system of by-molecules using cell-free biochemical reactions

Research Project

Project/Area Number 23360367
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionNagoya University

Principal Investigator

NAKANO Hideo  名古屋大学, 生命農学研究科, 教授 (00237348)

Co-Investigator(Kenkyū-buntansha) IWASAKI Yugo  名古屋大学, 大学院生命農学研究科, 准教授 (50273214)
KOJIMA Takaaki  名古屋大学, 大学院生命農学研究科, 助教 (40909080)
KOBAYASHI Isao  独立行政法人農業・食品産業技術総合研究機構, 食品総合研究所, 研究員 (70425552)
兒島 孝明  名古屋大学, 生命農学研究科, 助教 (40509080)
Project Period (FY) 2011-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2013: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2012: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2011: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000)
Keywords無細胞タンパク質合成系 / エマルジョン / タンパク質 / 酸化還元酵素 / RNAポリメラーゼ / 一分子PCR / マイクロビーズ / 酵素 / ハイスループットスクリーニング / ペルオキシダーゼ / スクリーニング / エマルジョンPCR / タンパク質工学 / セルラーゼ
Outline of Final Research Achievements

Creation of bio-macromolecules such as proteins, DNA and RNA is important technology both for bio-industries and bioscience, however, functional design of these molecules are generally impossible due to these complexities. In this research, we have developed a high-throughput library construction and screening system enabling giga-scale screening of such molecules using pL volume droplets in emulsion as reaction chamber for these molecules based on biochemical functions such as manganese peroxidase, horseradish peroxidase, and the substrate peptides and promoter sequences of transglutaminase and RNA polymerase, respectively.
In addition, hand-made device for making uniform droplets in emulsion formation in a short-time has been also developed to improve the screening efficiency.

Report

(5 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Annual Research Report
  • 2012 Annual Research Report
  • 2011 Annual Research Report
  • Research Products

    (28 results)

All 2015 2014 2013 2012 2011

All Journal Article (5 results) (of which Peer Reviewed: 4 results,  Acknowledgement Compliant: 2 results,  Open Access: 1 results) Presentation (23 results) (of which Invited: 2 results)

  • [Journal Article] Ultra-High-Throughput Screening of an <i>In Vitro</i>-Synthesized Horseradish Peroxidase Displayed on Microbeads Using Cell Sorter2015

    • Author(s)
      Bo Zhu, Takuro Mizoguchi, Takaaki Kojima, and Hideo Nakano
    • Journal Title

      PLOS ONE

      Volume: 未定

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Role of disulfide bond isomerase DsbC, calcium ions, and hemin in cell-free protein synthesis of active manganese peroxidase isolated from Phanerochaete chrysosporium.2014

    • Author(s)
      Ninomiya R, Zhu B, Kojima T, Iwasaki Y, Nakano H.
    • Journal Title

      J Biosci Bioeng.

      Volume: 117 Issue: 5 Pages: 652-657

    • DOI

      10.1016/j.jbiosc.2013.11.003

    • NAID

      110009823145

    • Related Report
      2014 Annual Research Report 2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] ビーズディスプレイ法を用いたDNA-転写因子間相互作用のハイスループット解析2014

    • Author(s)
      中野秀雄、兒島孝明
    • Journal Title

      バイオサイエンスとインダストリー

      Volume: 72 Pages: 306-308

    • NAID

      40020154978

    • Related Report
      2014 Annual Research Report
    • Acknowledgement Compliant
  • [Journal Article] 超微細生化学反応系」技術の最前線2014

    • Author(s)
      兒島孝明、中野秀雄
    • Journal Title

      化学と生物

      Volume: 52 Pages: 159-165

    • NAID

      130004808959

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Comprehensive Analysis of the DNA-Binding Specificity of an <i>Aspergillus nidulans</i> Transcription Factor, AmyR, Using a Bead Display System2012

    • Author(s)
      Wang, P., Kojima, T., Kobayashi, T., Nakano, H.
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 76 Issue: 6 Pages: 1128-1134

    • DOI

      10.1271/bbb.110949

    • NAID

      10030818030

    • ISSN
      0916-8451, 1347-6947
    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Presentation] In vitro transcription/translation in emulsion produced by a simple flow-focusing device.2015

    • Author(s)
      Marsel Murzabaev, Takuro Mizoguchi, Takaaki Kojima, Isao Kobayashi, Hideo Nakano.
    • Organizer
      249th ACS National Meeting & Exposition
    • Place of Presentation
      Denver, USA
    • Year and Date
      2015-03-22 – 2015-03-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] Bead display of heme enzymes for function-based high-throughput screening.2014

    • Author(s)
      Hideo Nakano, Eri Ota, Bo Zhu, Takaak Kojima.
    • Organizer
      Japan-Italy Joint Symposium.
    • Place of Presentation
      奈良
    • Year and Date
      2014-11-05 – 2014-11-06
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] In vitro synthesis of horseradish peroxidase with DNA binding tag and its fluorogenic assay for bead display-based ultra-high-throughput screening.2014

    • Author(s)
      Bo Zhu, Takuro Mizoguchi, Takaaki Kojima, Hideo Nakano.
    • Organizer
      Japan-Italy Joint Symposium
    • Place of Presentation
      奈良
    • Year and Date
      2014-11-05 – 2014-11-06
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] DNA結合タンパク質を用いたタンパク質の新規ビーズディスプレイ法の構築とトランスグルタミナーゼアッセイへの応用.2014

    • Author(s)
      溝口 琢郎, 兒島 孝明, 人見 清隆, 中野 秀雄.
    • Organizer
      日本農芸化学会中部支部 第171回例会
    • Place of Presentation
      名古屋
    • Year and Date
      2014-10-11
    • Related Report
      2014 Annual Research Report
  • [Presentation] DNA 結合タンパク質を用いたビーズディスプレイ法による糸状菌由来マンガンペルオキシダーゼのハイスループットスクリーニング系の開発2014

    • Author(s)
      太田 英里, 朱 博, 兒島 孝明, 中野秀雄
    • Organizer
      日本農芸化学会中部支部 第171回例会
    • Place of Presentation
      名古屋
    • Year and Date
      2014-10-11
    • Related Report
      2014 Annual Research Report
  • [Presentation] A simple flow-focusing device for high-throughput applications in emulsion.2014

    • Author(s)
      Marsel MURZABAEV, Takaaki KOJIMA, Isao KOBAYASHI, Hideo NAKANO
    • Organizer
      日本生物工学会2014年66回大会
    • Place of Presentation
      札幌
    • Year and Date
      2014-09-09 – 2014-09-11
    • Related Report
      2014 Annual Research Report
  • [Presentation] Cell-free Synthesis of Horseradish Peroxidase and Single-chain Lambda Cro Repressor Fusion Protein for Bead Display-based High-throughput Screening.2014

    • Author(s)
      Bo Zhu, Takuro Mizoguchi, Takaaki Kojima, Yugo Iwasaki and Hideo Nakano
    • Organizer
      The 7th International Congress on Biocatalysis 2014
    • Place of Presentation
      Hamburg
    • Year and Date
      2014-08-28 – 2014-09-01
    • Related Report
      2014 Annual Research Report
  • [Presentation] ビーズディスプレイ法を用いたマンガンペルオキシダーゼの酵素機能改変技術の構築2014

    • Author(s)
      太田 英里, 朱 博, 二宮 涼子, 兒島 孝明, 中野 秀雄
    • Organizer
      生物工学若手研究者の集い(若手会) 夏のセミナー2014
    • Place of Presentation
      神戸
    • Year and Date
      2014-07-12 – 2014-07-13
    • Related Report
      2014 Annual Research Report
  • [Presentation] Cell-free Synthesis of Horseradish Peroxidase and Single-chain Lambda Cro Repressor Fusion Protein for Bead Display-based High-throughput Screening.2014

    • Author(s)
      Bo Zhu, Takuro Mizoguchi, Takaaki Kojima, and Hideo Nakano.
    • Organizer
      71st Semi-annual Meeting of the Japanese Society of Enzyme Engineering
    • Place of Presentation
      福岡
    • Year and Date
      2014-04-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] DNA結合タンパク質を用いたタンパク質の新規ビーズディスプレイ法の開発.2013

    • Author(s)
      溝口琢郎, Murzabaev M., 小林功, 兒島孝明, 人見清隆, 中野秀雄: 平成25年9月,
    • Organizer
      日本生物工学会2013年65回大会,
    • Place of Presentation
      広島
    • Related Report
      2013 Annual Research Report
  • [Presentation] ビーズディスプレイ法を用いた糸状菌転写因子AmyR結合部位ゲノムワイドスクリーニング. ,2013

    • Author(s)
      7. 兒島孝明, 江崎俊文, 王ハン輝, 金丸京子, 小林哲夫, 中野秀雄
    • Organizer
      日本生物工学会2013年65回大会
    • Place of Presentation
      広島.
    • Related Report
      2013 Annual Research Report
  • [Presentation] エマルジョンPCRを用いたマイクロビーズ上へのDNAディスプレイ技術とその応用2013

    • Author(s)
      中野秀雄 兒島孝明
    • Organizer
      化学工学会第45回秋季大会
    • Place of Presentation
      岡山市
    • Related Report
      2013 Annual Research Report
  • [Presentation] Display of macromolecules on microbeads: a new platform for various screening methods.2013

    • Author(s)
      Nakano, H.
    • Organizer
      Enzyme Engineering XXII
    • Place of Presentation
      富山
    • Related Report
      2013 Annual Research Report
  • [Presentation] Cell-free synthesis of horseradish peroxidase and manganese peroxidase for bead display-based high-throughput screening.2013

    • Author(s)
      Zhu, B., Ninomiya, R., Kojima, T., Iwasaki, Y., Nakano, H.
    • Organizer
      日本農芸化学会中部支部第168回例会,
    • Place of Presentation
      名古屋
    • Related Report
      2013 Annual Research Report
  • [Presentation] : DNA結合タンパク質を用いたタンパク質の新規ビーズディスプレイ法の開発とトランスグルタミナーゼアッセイへの応用.2013

    • Author(s)
      溝口琢郎, Murzabaev, M., 小林功, 兒島孝明, 人見清隆, 中野秀雄
    • Organizer
      日本農芸化学会中部支部第168回例会,
    • Place of Presentation
      名古屋
    • Related Report
      2013 Annual Research Report
  • [Presentation] Uniform emulsion for biochemical applications using inexpensive flow-focusing device.2013

    • Author(s)
      Murzabaev, M., Kojima, T., Kobayashi, I., Nakano, H.
    • Organizer
      日本農芸化学会中部支部第168回例会,
    • Place of Presentation
      名古屋
    • Related Report
      2013 Annual Research Report
  • [Presentation] 無細胞タンパク質合成系を用いたライブラリー構築技術の開発と応用2012

    • Author(s)
      中野秀雄、兒島孝明
    • Organizer
      日本農芸化学会2012大会
    • Place of Presentation
      京都(招待講演)
    • Year and Date
      2012-03-24
    • Related Report
      2011 Annual Research Report
  • [Presentation] Development of bead display system for molecular evolution of cellulases2012

    • Author(s)
      朱博, 森翔也、松田秀樹、兒島孝明、中野秀雄
    • Organizer
      日本農芸化学会2012大会
    • Place of Presentation
      京都
    • Year and Date
      2012-03-23
    • Related Report
      2011 Annual Research Report
  • [Presentation] Comprehensive analysis of transcription factor binding site using bead display of DNA.2012

    • Author(s)
      Nakano H
    • Organizer
      The 12th Japan-China-Korea Joint Symposium on Enzyme Engineering,
    • Place of Presentation
      金沢
    • Related Report
      2012 Annual Research Report
  • [Presentation] Development of bead display system for high-throughput screening of cellulases using emulsion PCR and cell-free protein synthesis system.2012

    • Author(s)
      Zhu B
    • Organizer
      The 12th Japan-China-Korea Joint Symposium on Enzyme Engineering,
    • Place of Presentation
      金沢
    • Related Report
      2012 Annual Research Report
  • [Presentation] ビーズディスプレイ法を用いた糸状菌由来リグニン分解酵素のハイスループットスクリーニング系の構築2012

    • Author(s)
      二宮涼子
    • Organizer
      日本生物工学会2012年大会
    • Place of Presentation
      神戸
    • Related Report
      2012 Annual Research Report
  • [Presentation] ビーズディスプレイ法を用いた糸状菌由来リグニン分解酵素のハイスループットスクリーニング系の構築2012

    • Author(s)
      二宮涼子
    • Organizer
      日本農芸化学会中部支部第165回例会
    • Place of Presentation
      名古屋
    • Related Report
      2012 Annual Research Report
  • [Presentation] エマルジョンPCRを用いた分子間相互作用ハイスループットスクリーニング系2011

    • Author(s)
      中野秀雄、兒島孝明
    • Organizer
      平成23年度生物工学技術セミナー
    • Place of Presentation
      東京(招待講演)
    • Year and Date
      2011-11-30
    • Related Report
      2011 Annual Research Report

URL: 

Published: 2011-04-06   Modified: 2019-07-29  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi