Preparation of ochre suppressor tRNA and its use for expansion of the genetic code

• Project/Area Number: 19550161
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Chemistry related to living body
• Research Institution: Gifu University
• Principal Investigator: NISHIKAWA Kazuya Gifu University, 工学部, 教授 (60109262)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2008: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2007: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: バイオテクノロジー / 核酸・蛋白質・糖化学 / 生体認識・機能化学 / 遺伝暗号 / サプレッサー

Research Abstract

遺伝暗号表の拡張を目指して、UAA (Ochre)コドンに対応できるサプレッサーtRNA を調製し、生体外タンパク質合成系においてOchre コドン選択的に非天然アミノ酸をタンパク質に導入する方法を開発した。分子整形技術により調製したtRNA (ΨΨA)は期待通りのOchre サプレッサーとして機能したが、意外にも対照のtRNA (UΨA)も同様の挙動を示した。この結果は「ウォブル説」の再吟味も含め、さらなる検討の必要性を示唆する。

Research Products

• [Journal Article] Site-selective post-translational modification of proteins using an unnatural amino acid (2007)
  • Author(s): S. Ohno, M. Matsui, T. Yokogawa, M. Nakamura, T. Hosoya, T. Hiramatsu, M. Suzuki, N. Hayashi, and K. Nishikawa
  • Journal Title: 3-azidotyrosine. J. Biochem 141 Pages: 335-343
  • Peer Reviewed
• [Journal Article] Structural basis for recognition of cognate tRNA by tyrosyl-tRNA synthetase from three kingdoms (2007)
  • Author(s): M. Tsunoda, Y. Kusakabe, N. Tanaka, S. Ohno, M. Nakamura, T. Senda, T. Moriguchi, N. Asai, M. Sekine, T. Yokogawa, K. Nishikawa, and K. T. Nakamura
  • Journal Title: Nucleic Acids Res 35 Pages: 4289-4300
  • Peer Reviewed
• [Journal Article] Extremely thermophilic translation system in the common ancestor commonote : ancestral mutants of glycyl-tRNA synthetase from the extreme thermophile Thermus thermophilus (2007)
  • Author(s): H. Shimizu, S.-i. Yokobori, T. Ohkuri, T. Yokogawa, K. Nishikawa and A. Yamagishi
  • Journal Title: J. Mol. Biol 369 Pages: 1060-1069
  • Peer Reviewed
• [Journal Article] Site-selective post-translational modification of proteins using an unnatural amino acid, 3-azidotyrosine. (2007)
  • Author(s): S. Ohno, M. Matsui, T. Yokogawa, M. Nakamura, T. Hosoya, T. Hiramatsu, M. Suzuki, N. Hayashi, and K. Nishikawa
  • Journal Title: J. Biochem. 141 Pages: 335-343
  • NAID: 10020100554
  • Peer Reviewed
• [Journal Article] Structural basis for recognition of cognate tRNA by tyrosyl-tRNA synthetase from three kingdoms. (2007)
  • Author(s): M. Tsunoda, Y. Kusakabe, N. Tanaka, S. Ohno, M. Nakamura, T. Senda, T. Moriguchi, N. Asai, M. Sekine, T. Yokogawa, K. Nishikawa, and K.T.Nakamura
  • Journal Title: Nucleic Acids Res. 35 Pages: 4289-4300
  • Peer Reviewed
• [Journal Article] Extremely thermophilic translation system in the common ancestor commonote: ancestral mutants of glycyl-tRNA synthetase from the extreme thermophile Thermw thermophilus. (2007)
  • Author(s): H. Shimizu, S.-i. Yokobori, T. Ohkuri, T. Yokogawa, K. Nishikawa and A. Yamagishi
  • Journal Title: J. Mol. Biol. 369 Pages: 1060-1069
  • Peer Reviewed