Analysis of DNA replication initiation based on controlling technique of single DNA molecules

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K07280
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Gunma University
• Principal Investigator: KATSURA Shinji 群馬大学, 大学院理工学府, 教授 (10260598)
• Co-Investigator(Kenkyū-buntansha): 大重 真彦 群馬大学, 大学院理工学府, 准教授 (00451716)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: DNAポリメラーゼ / DNA複製 / DNA超らせん構造

Outline of Final Research Achievements

Single molecule analysis of DNA elongation rates of several DNA polymerases have been carried out. This study demonstrated that processive polymerase, T7 DNA polymerase synthesized with 144 bases/s in elongation rate, and the pocessivity was also determined as 11.1kbases. On the other hand, any pauses in polymerase reactions have not been observed in the case of a distributive polymerase DNA polymerase I, probably due to too short processivity less than a resolution limit of an optical microscope.
We also succeeded in development of fluorescent observation system with supercoil induction, which permitted real time observation of DNA behavior during supercoil induction.

Free Research Field

生物化学工学

Academic Significance and Societal Importance of the Research Achievements

DNAポリメラーゼの挙動を1分子レベルで解析する方法を確立した。この方法は他のポリメラーゼのキャラクタリゼーションにも適用できると考えられ、DNAポリメラーゼの分類・機能分担の解析に応用できると考えている。
また、併せてDNAへ超らせん構造を導入しながら蛍光解析を行うシステムを開発したので、超らせん構造がDNA代謝酵素に与えている影響を解析できると考えられる。