Novel generic technology for genome synthesis derived from Horizontal gene transfer in nature

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K06196
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43060:System genome science-related
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: KANEKO Shinya 東京工業大学, 生命理工学院, 助教 (10399694)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: ゲノム合成 / コンティグDNA / 枯草菌 / 応用微生物 / 形質転換 / 核酸 / ゲノム / バイオテクノロジー

Outline of Final Research Achievements

Handling of DNA fragments comprising over several dozens of kb is difficult, because of their fragility when they are in solution. To avoid physical shearing in vitro, assembly of contig DNA fragments have been accomplished within Bacillus subtilis cell using one step protoplast transformation method. Moreover, to avoid physical shearing in the purification step, we demonstrated that large DNA plasmid over 50 kb in size can be introduced into Escherichia coli, or Saccharomyces cerevisiae using the lysate of donor B. subtilis cells. Conjugation protocol has also been established for transfer of large-sized DNA from E. coli to
eukaryotic microorganisms (S. cerevisiae). Although transfer to animal culture cell remains, our designed horizontal transfer methods is a versatile procedure for delivery of large-sized DNAs into prokaryotic and eukaryotic microorganisms, which would be a basic platform in the synthetic genome area.

Free Research Field

応用ゲノム工学

Academic Significance and Societal Importance of the Research Achievements

生物の設計図であるゲノム配列を簡単に解読できる時代に突入した。次の段階としてゲノム（長鎖DNA）を設計・合成し、細胞導入する研究が世界各国で進行している。しかしあまり認識されていないが、DNAは長くなればそれだけ物理的な衝撃に弱く扱いが難しくなる。本研究の成果は簡便に「長鎖DNAを合成する」だけでなく、合成後の「長鎖DNAを如何に目的の宿主に導入するか」を解決する上で重要な基盤技術を提供する。