DNA uptake mechanism of E. coli transformation

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K05782
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 38020:Applied microbiology-related
• Research Institution: Rikkyo University
• Principal Investigator: Nozaki Shingo 立教大学, 理学部, 助教 (70725481)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 大腸菌 / 形質転換 / トランスフォーメーション / コンピテントセル

Outline of Final Research Achievements

Factors involved in E. coli transformation were screened and deletion of envZ gene was found to increase transformation efficiency. This increase was due to the increase in expression level of OmpA, one of the outer membrane proteins. It became clear that OmpA plays an important role in transformation. In addition, as a result of challenge to develop a simple transformation method, it was found that competent cells were prepared from 0.05 mL culture in a micro test tube. It is expected that transformation can be performed more easily by being able to prepare competent cells from such a small amount of E. coli culture.

Free Research Field

応用微生物学

Academic Significance and Societal Importance of the Research Achievements

大腸菌の形質転換はDNA組換え技術において最も基本的な操作の一つである。その機構が解明されることで、形質転換効率がより向上した高性能で使い勝手の良い宿主大腸菌株や新たな形質転換法を生み出す基盤となる。また、DNAのような巨大分子の取り込み機構の解明は、細胞膜を透過しづらい抗菌剤を細胞内に浸透させるための大きな手がかりとなる。これは、多剤耐性菌が社会的な問題となっている現代において、抗菌剤を有効に活用する新たな手段の開発にもつながるものである。