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Molecular mechanism of Cryptococcus liquefaciens N6 strain

Research Project

Project/Area Number 15K14434
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field System genome science
Research InstitutionTokyo Institute of Technology

Principal Investigator

Iwasaki Hiroshi  東京工業大学, 科学技術創成研究院, 教授 (60232659)

Research Collaborator ITO Takehiko  東京工業大学, 生命理工学院, 教授 (90501106)
Palihati Maierdan  東京工業大学, 生命理工学院, 大学院生
Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
KeywordsCryptococcus酵母 / 形質転換 / 分子生物学 / 銅耐性 / 分子生物学ツール / モデル生物 / Cryptococcus / 酵母 / ゲノム編集 / ウラシル要求性 / 相同組換え / Cryptococcus 酵母 / 銅イオン / プラスミド
Outline of Final Research Achievements

Cryptococcus liquefaciens N6 is a unique strain of yeast that shows resistance to copper toxicity. To elucidate the mechanism underlying this phenomenon, we aimed to establish molecular biology tools in C. liquefaciens. For this, we first isolated ura- mutants by using 5-FOA as a positive selection marker. Two ura- mutants were found to have the same nonsense mutation in the URA5 gene by whole genome sequencing. Next, we established conditions for genetic manipulation of one ura5 mutant by electroporation, resulting in ura--to-ura+ transformation. We then attempted gene disruption of URA5 through a double crossover event with a targeting vector containing the NAT resistance gene. Although we obtained a considerable number of NAT resistant transformants, the NAT resistance gene was found to have integrated in an ectopic location in all cases. We speculate that gene disruption by this approach was unsuccessful due to intrinsically inefficient homologous recombination in C. liquefaciens.

Report

(3 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • Research Products

    (9 results)

All 2017 2016 Other

All Journal Article (3 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 3 results,  Open Access: 3 results,  Acknowledgement Compliant: 2 results) Presentation (4 results) (of which Int'l Joint Research: 2 results,  Invited: 4 results) Book (1 results) Remarks (1 results)

  • [Journal Article] In vitro site-specific recombination mediated by the tyrosine recombinase XerA of Thermoplasma acidophilum.2017

    • Author(s)
      Jo M, Murayama Y, Tsutsui Y and Iwasaki H.
    • Journal Title

      Genes to Cells

      Volume: 印刷中 Issue: 7 Pages: 646-661

    • DOI

      10.1111/gtc.12503

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Holliday junction resolvases mediate chloroplast nucleoid segregation2017

    • Author(s)
      Kobayashi Y, Misumi O, Odahara M, Ishibashi K, Hirono M, Hidaka K, Endo M, Sugiyama H, Iwasaki H, Kuroiwa T, Shikanai T, Nishimura Y.
    • Journal Title

      Science

      Volume: 356 Issue: 6338 Pages: 631-634

    • DOI

      10.1126/science.aan0038

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] 1.The Differentiated and Conserved Roles of Swi5-Sfr1 in Homologous Recombination.2017

    • Author(s)
      Argunhan B, Murayama Y and Iwasaki H.
    • Journal Title

      FEBS letter

      Volume: - Issue: 14 Pages: 2035-2047

    • DOI

      10.1002/1873-3468.12656

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] DNA鎖交換反応のリアルタイム解析2016

    • Author(s)
      岩崎博史
    • Organizer
      第89回日本生化学会大会:シンポジウム「ゲノムの維持と継承を制御するタンパク質と核酸の相互作用」
    • Place of Presentation
      東北大学、宮城
    • Year and Date
      2016-09-25
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 分裂酵母の接合型変換の分子機構2016

    • Author(s)
      岩崎博史
    • Organizer
      第68回日本細胞生物学会大会:シンポジウム「染色体諸機能の連携と機能統合体としての理解」
    • Place of Presentation
      京都テルサ、京都
    • Year and Date
      2016-06-15
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] Real-time analysis of Rad51-mediated DNA strand exchange activation by Swi5-Sfr1 complex2016

    • Author(s)
      Hiroshi Iwasaki
    • Organizer
      Abcam conference: Mechanisms of Recombination
    • Place of Presentation
      Alicante, Spain
    • Year and Date
      2016-05-16
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Post-synaptic roles of Swi5-Sfr1 in DNA strand exchange2016

    • Author(s)
      Hiroshi Iwasaki
    • Organizer
      Protein-DNA complexes in homologous recombination at Szent-Gyoergyi Day of the hungarian protein and medicinal community (MedinProt)
    • Place of Presentation
      Hungarian Academy of Sciences, Budapest, Hungary
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Book] 池上彰か聞いてわかった生命の仕組み2016

    • Author(s)
      池上彰、岩崎博史、田口英樹
    • Total Pages
      246
    • Publisher
      朝日新聞出版
    • Related Report
      2016 Annual Research Report
  • [Remarks]

    • URL

      http://www.iwasakilab.bio.titech.ac.jp

    • Related Report
      2016 Annual Research Report

URL: 

Published: 2015-04-16   Modified: 2018-03-22  

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