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Elucidation of the mechanism of translation enhancement in Escherichia coli by exogenous nucleotide sequences

Research Project

Project/Area Number 19K15809
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 38060:Applied molecular and cellular biology-related
Research InstitutionThe University of Tokyo

Principal Investigator

Kondo Tomo  東京大学, 大学院総合文化研究科, 助教 (50728293)

Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords大腸菌 / 物質生産 / 翻訳促進 / 遺伝子発現 / 翻訳 / 画像解析 / タンパク質発現 / タンパク質翻訳 / 細胞性粘菌
Outline of Research at the Start

大腸菌を用いた異種タンパク質の生産は,研究室レベルに留まらず産業レベルでも基盤的な技術として利用される。例えばインスリンの製造をはじめとして,抗体を用いるバイオ医薬品などの製造にも近年では利用され,現在でも用途は拡大している。製造コストを抑えながら大量に物質生産ができることが,社会にとって究極的に有益かつ重要であるが,その技術開発は頭打ちの状態である。
そこで本研究では,研究代表者らが最近見出した「大腸菌のタンパク質翻訳が外来の塩基配列によって促進される現象」を基にしたタンパク質生産力の強化に関する技術開発とその法則性の解明に取り組む。

Outline of Final Research Achievements

In this study, I sought to elucidate the mechanism of the phenomenon of translation enhancement by foreign nucleotide sequences in E. coli, which was discovered earlier by us. First, I focused on the amount of transcripts of genes under the influence of TED, and examined the amount of mRNA of reporter genes. I found that the amount of mRNA of the reporter gene was several times higher than that of the control. Furthermore, I investigated a method to reproduce TED in vitro and succeeded in reproducing TED by using purified molecules such as ribosomes from E. coli. Importantly, TED was observed even when the same amount of mRNA as the control was used in the reaction. Taken together, I hypothesized that the mechanisms of TED in cells include 1) quantitative change of mRNA by suppression of its degradation, and 2) acceleration of translation initiation by ribosomes.

Academic Significance and Societal Importance of the Research Achievements

大腸菌を用いた異種タンパク質の生産は,基礎研究のみならず産業レベルでも基盤的な技術である。我々が見出したTEDと今回明らかにしたそのメカニズムは,これまで生産技術改良が頭打ちになっていたところに,増産に向けた開発の新しい端緒と視点を与えるものである。基礎研究でも簡単に導入可能である。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (3 results)

All 2020 2019

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (1 results)

  • [Journal Article] Strategies for enhancing gene expression in Escherichia coli2020

    • Author(s)
      Kondo Tomo、Yumura Shigehiko
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 104 Issue: 9 Pages: 3825-3834

    • DOI

      10.1007/s00253-020-10430-4

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] An improved molecular tool for screening bacterial colonies using GFP expression enhanced by a Dictyostelium sequence2020

    • Author(s)
      Kondo Tomo、Yumura Shigehiko
    • Journal Title

      BioTechniques

      Volume: 68 Issue: 2 Pages: 91-95

    • DOI

      10.2144/btn-2019-0127

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 外来塩基配列による翻訳促進効果を利用した大腸菌タンパク質発現系の改良2019

    • Author(s)
      近藤興
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Research-status Report

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Published: 2019-04-18   Modified: 2023-01-30  

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