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

Research Project

Project/Area Number 17K07280
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Molecular biology
Research InstitutionGunma 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
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
KeywordsDNAポリメラーゼ / DNA複製 / DNA超らせん構造 / DNA超らせん / 1分子観察 / DNA合成 / 1分子観察 / 超らせん
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.

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。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (3 results)

All 2019 2018 2017

All Presentation (3 results)

  • [Presentation] 1分子解析のためのマイクロ流路内におけるDNA分子の片端固定化法2019

    • Author(s)
      青木凌太、勝見匡 、高橋俊介 、川崎祥平 、栗田弘史 、水野武 、水野彰、大重真彦、桂進司
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 発光酵素ルシフェラーゼによるDNA合成反応の検出2018

    • Author(s)
      栗本雅之、小野稜平、手塚大輔、樋山みやび、板橋英之、大重真彦、桂進司
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] SSB-YFPを用いたDNAポリメラ-ゼの1分子解析2017

    • Author(s)
      勝見 匡、 栗本雅之、 高橋俊介、 川崎祥平、 栗田弘史、 松浦俊一 、水野 彰、大重真彦、桂 進司
    • Organizer
      生物科学系学会合同年次大会
    • Related Report
      2017 Research-status Report

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Published: 2017-04-28   Modified: 2021-02-19  

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