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2013 Fiscal Year Final Research Report

Creation of functional peptides recognizing a low-molecular compound by evolutionary molecular engineering

Research Project

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Project/Area Number 23510253
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Living organism molecular science
Research InstitutionSaitama University

Principal Investigator

NEMOTO Naoto  埼玉大学, 理工学研究科, 准教授 (60509727)

Project Period (FY) 2011 – 2013
Keywords進化分子工学 / ペプチド / アプタマー / アミノ基認識 / 分子間相互作用 / 試験管内淘汰 / cDNA display / mRNA display
Research Abstract

We have succeeded to improve the preparation of cDNA display molecule easily and efficiently with a one-pot preparational manner. Furthermore, we demonstrated the biotin binding capacity of streptavidin magnetic beads decreased by mRNA-binding ribosomes, resulted in the low yield of cDNA display molecules. Secondly, we developed an easy and rapid molecular-protein interaction analysis using a cell-free translation and a novel puromycin-linker by a pull-down manner. Finally, by using these techniques, in vitro selection using 30 amino acids random library containing cysteine against amino group on the solid-phase was performed. As a result, an amino group-binding peptide aptamer containing two disulfide bonds which have a unique structure has explored. This shows that a peptide has capability to recognize small molecules with a low-molecular weight. In the future, peptides will be more and more applicable for nanotechnology as a recogniton material of nano sturcture.

  • Research Products

    (22 results)

All 2014 2013 2012 2011 Other

All Journal Article (7 results) (of which Peer Reviewed: 7 results) Presentation (10 results) Book (4 results) Remarks (1 results)

  • [Journal Article] Amino group binding peptide aptamers with double disulphide-bridged loops selected by in vitro selection using cDNA display2014

    • Author(s)
      Y. Mochizuki, K. Nishigaki, N. Nemoto
    • Journal Title

      Chem Commun

      Volume: 50 Pages: 5608-5610

    • Peer Reviewed
  • [Journal Article] cDNA display による分子デザイン- mRNA display(in vitro virus)からcDNA display へ-2013

    • Author(s)
      根本直人、望月佑樹、上野真吾
    • Journal Title

      生物物理

      Volume: 53 Pages: 250-253

    • Peer Reviewed
  • [Journal Article] Increasing the library size in cDNA display by optimizing purification procedures2013

    • Author(s)
      Y. Mochizuki, S. Kumachi, K. Nishigaki, N. Nemoto
    • Journal Title

      Biol Proced Online

      Volume: 15 Pages: 7

    • Peer Reviewed
  • [Journal Article] A pull-down method with a biotinylated bait protein prepared by cell-free translation using a puromycin-linker2013

    • Author(s)
      Y. Mochizuki, F. Kohno, K. Nishigaki, N. Nemoto
    • Journal Title

      Anal. Biochem

      Volume: 434 Pages: 93-95

    • Peer Reviewed
  • [Journal Article] Antagonistic effect of disulfide-rich peptide aptamers selected by cDNA display on interleukin-6-dependent cell proliferation2012

    • Author(s)
      N. Nemoto, C. Tsutsui, J. Yamaguchi, S. Ueno, M. Machida, T. Kobayashi, T. Sakai
    • Journal Title

      Biochem Biophys Res Commun

      Volume: 421 Pages: 129-133

    • Peer Reviewed
  • [Journal Article] In vitro Selection of Peptide Aptamers against Acetylcholine-binding Protein (AChBP) from Disulfide-rich Peptide Library by cDNA Display2012

    • Author(s)
      N. Nemoto, S. Kimura, S. Kumachi, W. Cai, M. Suzuki, K. Nishigaki, and T. Kubo
    • Journal Title

      Peptide Science 2011: K. Sakaguchi (Ed.)

      Pages: 89-90

    • Peer Reviewed
  • [Journal Article] One-pot preparation of mRNA/cDNA display by a novel and versatile puromycin-linker DNA2011

    • Author(s)
      Y. Mochizuki , M. Biyani , S. T. -Ueno , M. Suzuki , K. Nishigaki , Y. Husimi , and N. Nemoto
    • Journal Title

      ACS Comb. Sci

      Volume: 13 Pages: 478-485

    • Peer Reviewed
  • [Presentation] ペプチドで広がる医療・診断技術 - ペプチドアプタマーを中心にして -2013

    • Author(s)
      根本直人
    • Organizer
      第4回 「医工連携によるイノベーション創出研究会」
    • Place of Presentation
      さいたま市
    • Year and Date
      2013-12-11
  • [Presentation] cDNA display 法を用いた TNFα に対するシステインリッチペプチドライブラリの試験管内淘汰実験2013

    • Author(s)
      澤田 貴宏,望月 佑樹、鈴木 美穂,西垣功一、根本 直人
    • Organizer
      第36回日本分子生物学会年会
    • Place of Presentation
      神戸
    • Year and Date
      2013-12-05
  • [Presentation] 標的分子の多様性を拡張するペプチドアプタマー取得ための cDNA display 法2013

    • Author(s)
      根本直人
    • Organizer
      Biotech 2013
    • Place of Presentation
      東京(東京ビックサイト)
    • Year and Date
      2013-05-09
  • [Presentation] Easy-to-use and Rapid Molecular Interaction Analysis by Combination of Pull-down method with mRNA Display2012

    • Author(s)
      F. Kono, Y. Mochizuki, M. Suzuki, K. Nishigaki, N. Nemoto
    • Organizer
      The 35th Annual Meeting of the Molecular Biology Society of Japan
    • Place of Presentation
      Fukuoka
    • Year and Date
      2012-12-12
  • [Presentation] Optimization of In Vitro Protein Selection using mRNA/cDNA Display Method to Search Functional Peptides and Proteins Rapidly2012

    • Author(s)
      Y. Mochizuki, N. Nemoto
    • Organizer
      Protein & Peptide Conference 2012
    • Place of Presentation
      Beijing, China
    • Year and Date
      2012-03-25
  • [Presentation] In vitro evolution of peptide aptamers against survivin from a novel three-finger scaffold library by cDNA display2011

    • Author(s)
      Y. Yotsumoto,Y. Mochizuki,M. Suzuki,K. Nishigaki,N. Nemoto
    • Organizer
      第34回日本分子生物学会年会(MBSJ2011)
    • Place of Presentation
      横浜
    • Year and Date
      2011-12-13
  • [Presentation] cDNA display 法によるジスルフィド結合を複数含むペプチドライブラリからのアセチルコリン結合タンパク質結合ペプチドアプタマーの試験管内淘汰2011

    • Author(s)
      根本直人、木村真之介、熊地重文、Cai Weiyan、鈴木美穂、西垣功一、久保 泰
    • Organizer
      第48回ペプチド討論会
    • Place of Presentation
      札幌
    • Year and Date
      2011-09-29
  • [Presentation] タンパク/ペプチドの簡易で迅速な精製・ビオチン化法2011

    • Author(s)
      根本直人
    • Organizer
      イノベーション・ジャパン2011「新技術説明会」
    • Place of Presentation
      東京
    • Year and Date
      2011-09-21
  • [Presentation]2011

    • Author(s)
      T. Fukushima, W. Cai, M. Suzuki, K. Nishigaki, T. Kubo, N. Nemoto
    • Organizer
      第49回日本物理学会年会
    • Place of Presentation
      姫路
    • Year and Date
      2011-09-17
  • [Presentation] cDNA display 法による新機能分子の創出2011

    • Author(s)
      根本直人
    • Organizer
      理研シンポジウム 第6回「バイオものづくり」シンポジウム
    • Place of Presentation
      和光
    • Year and Date
      2011-05-10
  • [Book] 遺伝子型-表現型対応付け技術,『進化分子工学-高速分子進化によるタンパク質・核酸の開発-』 第2編 第3章2013

    • Author(s)
      根本直人
    • Total Pages
      161-170
    • Publisher
      (株)エヌ・ティー・エス
  • [Book] Evolutionary Molecular Engineering to Efficiently Direct in vitro Protein Synthesis, CELL-FREE PROTEIN SYNTHESIS (Edited by Manish Biyani)2012

    • Author(s)
      M. Biyani, M. Biyani, N. Nemoto, Y. Husimi
    • Total Pages
      51-62
    • Publisher
      InTech
  • [Book] Evolution of Disulfide-Rich Peptide Aptamers Using cDNA Display, Methods in Mol. Biol2012

    • Author(s)
      Y. Mochizuki, N. Nemoto
    • Total Pages
      805, 237-250
    • Publisher
      Springer
  • [Book] cDNA Display : Rapid Stabilization of mRNA Display, Methods in Mol. Biol2012

    • Author(s)
      S. Ueno, N. Nemoto
    • Total Pages
      805, 113-135
    • Publisher
      Springer
  • [Remarks] ホームページ等

    • URL

      http://park.saitama-u.ac.jp/~nemoto/

URL: 

Published: 2015-07-16  

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