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

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K15809
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 38060:Applied molecular and cellular biology-related
• Research Institution: The 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
• Keywords: 大腸菌 / 物質生産 / 翻訳促進

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.

Free Research Field

細胞生物学

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。