Reconstitution of protein synthesis system of mammals

2009 Fiscal Year Final Research Report

• Project/Area Number: 19580113
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: University of Hyogo (2008-2009); The Institute of Physical and Chemical Research (2007)
• Principal Investigator: IMATAKA Hiroaki University of Hyogo, 大学院・工学研究科, 教授 (50201942)
• Project Period (FY): 2007 – 2009
• Keywords: タンパク質合成 / 無細胞システム / 翻訳 / 翻訳因子

Research Abstract

We purified mammalian translation initiation factors : eIF1, eIF1A, eIF2, eIF2B, eIF3, eIF4E, eIF4A, eIF4B, eIF4G, eIF5, eIF5B, elongation factors : eEF1A eEF1B・eEF2, termination factors : eRF1、eRF3, aminoacyl tRNA, and ribosome subunits (40S, 60S). By mixing all these factors, we successfully established a reconstituted protein synthesis system derived from mammals.

Research Products

• [Journal Article] N-terminally truncated GADD34 proteins are convenient translation enhancers in a human cell-derived in vitro protein synthesis system. (2010)
  • Author(s): Satoshi Mikami, Tominari Kobayashi, Kodai Machida, Mamiko Masutani, Shigeyuki Yokoyama, Hiroaki Imataka
  • Journal Title: Biotechnology letters DOI: 10. 1007/s10529-010-0251-7
  • Peer Reviewed
• [Journal Article] A human cell-derived in vitro coupled transcription/translation system (2008)
  • Author(s): Satoshi Mikami, Tominari Kobayashi, Mamiko Masutani, Shigeyuki Yokoyama, Hiroaki Imataka
  • Journal Title: optimized for production of recombinant proteins Protein Expr. Purif 62 Pages: 190-198
  • Peer Reviewed
• [Journal Article] An improved cell-free system for picornavirus synthesis (2007)
  • Author(s): Tominari Kobayashi, Satoshi Mikami, Shigeyuki Yokoyama, Hiroaki Imataka
  • Journal Title: J. Virol. Methods 142 Pages: 182-188
  • Peer Reviewed
• [Presentation] 試験菅内ウイルス粒子合成技術の開発 (2010)
  • Author(s): 今高寛晃
  • Organizer: 第14回けいはんなシーズフォーラム
  • Place of Presentation: 中ノ島センタービル、大阪市
  • Year and Date: 2010-01-26
• [Presentation] A novel human in vitro translation system for glycoprotein expression (2009)
  • Author(s): K. Vattem, S. Mikami, K. Maas, E. Hommema, A. Morgan, M. Schofield, B. Webb, A. Deshpande, H. Imataka
  • Organizer: 第8回国際プロテオミクス会議
  • Place of Presentation: トロント
  • Year and Date: 2009-09-28
• [Presentation] Human Cell-Derived in vitro Coupled Transcription/Translation Systems (2009)
  • Author(s): S Mikami, K Machida S Yokoyama, H Imataka
  • Organizer: 第8回国際プロテオミクス会議
  • Place of Presentation: トロント
  • Year and Date: 2009-09-28
• [Presentation] 動物細胞由来セルフリータンパク質合成系の確立 (2008)
  • Author(s): 三上暁、小林富成、舛谷真美子、横山茂之、今高寛晃
  • Organizer: 2008年度日本農芸化学会
  • Place of Presentation: 名城大学、名古屋
  • Year and Date: 20080327-20080329
• [Presentation] 強力なヒト細胞由来セルフリータンパク質合成システムの開発 (2007)
  • Author(s): 三上暁、小林富成、横山茂之、今高寛晃
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: パシフィコ横浜、横浜市
  • Year and Date: 20071211-20071215
• [Book] Cell-free protein synthesis systems with extracts from cultured human cells. In: Endo, Y., Takai, K. and Ueda, T. (Eds.), Methods in Molecular Biology 607, Cell-free protein production: Methods and Protocols. (2010)
  • Author(s): Satoshi Mikami, Tominari Kobayashi, Hiroaki Imataka
  • Total Pages: 43-52
  • Publisher: Human Press
• [Book] 大腸菌抽出液を用いた無細胞タンパク質合成系, タンパク質科学実験法1, タンパク質をつくるー抽出・精製と合成(長谷俊治、高尾敏文、高木淳一、編集) (2008)
  • Author(s): 横山茂之, 今高寛晃
  • Total Pages: 135-138
  • Publisher: 化学同人
• [Book] mrnaの調製ほ乳動物細胞由来in vitro翻訳システムRNA実験ノート (2007)
  • Author(s): 今高寛晃
  • Publisher: 羊土社
• [Book] 翻訳開始の制御
  • Author(s): 今高寛晃、三上暁
  • Total Pages: 2213-2218
  • Publisher: 蛋白質核酸酵素
• [Book] ヒト細胞由来無細胞タンパク質合成システムの魅力
  • Author(s): 今高寛晃、三上暁
  • Total Pages: 303-307
  • Publisher: 生化学