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Investigation of microbial systems using NGS driven powerful forward genetics.

Research Project

Project/Area Number 19K06627
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43060:System genome science-related
Research InstitutionNagoya University

Principal Investigator

Ihara Kunio  名古屋大学, 遺伝子実験施設, 准教授 (90223297)

Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2019: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywordsゲノム / 古細菌 / システム解析 / 遺伝子 / ハイスループット / 変異 / 表現型 / 選択圧 / 高度好塩性古細菌 / ゲノム解析 / UV変異 / ゲノム多様性 / 化学変異原 / 光回復 / ゲノムシーケンス / 次世代ゲノムライブラリー作成 / 好塩性古細菌 / UV照射 / 突然変異 / 微生物 / 好アルカリ性古細菌 / 走性 / 次世代シーケンス / トランスポゼース / 磁気ビーズ / ハイスループット化 / 遺伝学 / システム
Outline of Research at the Start

様々な生物のゲノムが大量に読まれる時代が到来した。しかし、ゲノム中の遺伝子 (ORF)の機能は分かっていないものが多く、ゲノムから生命を理解する際の大きな課題の1つとなっている。次世代シーケンサー(NGS)解析で強化された順方向遺伝学は、ゲノム中の遺伝子 (ORF) を生物機能システムの構成要素として理解する強力な手法である。このパワフルな順遺伝学を微生物の走性システムに適用し、システムに関係するORFを推定すると同時に、新規に得られる関連遺伝子数から、走性システムのサイズ(関係する遺伝子数)を見積もる手法を構築する。

Outline of Final Research Achievements

To achieve our purpose, we have attempted to construct a random mutation library with a sufficient number of mutations. In the process, we confirmed through mass genome analysis that mutagenic treatments such as UV irradiation or EMS mutagen
treatment, which had been thought to efficiently introduce mutations, did not introduce so many mutations even under low survival rate conditions (up to an average of 0.5 mutations/genome in the case of UV mutagenesis). Even with this low mutation rate, it was found that selection under some stress condition like phosphate deficiency increased the number of mutations introduced in the genome. In addition, mass genome analysis of more than 1,000 strains of one type of microorganism revealed the existence of potential genomic diversity in cryopreserved strains.

Academic Significance and Societal Importance of the Research Achievements

学術的には、これまで遺伝子変異導入に使われてきたUV変異による突然変異導入効率を、1000株程度のゲノム解析を行うことで決定し、高度好塩性好アルカリ性古細菌では、最大でも平均0.5個/ゲノム程度であることを決めたことと、凍結保存した株においてもかなりのゲノム多様性が存在することを明らかにしたことが、今後の知見として重要であると考える。社会的には、UVなどの変異原に対する生物の処理方法(完全に修復するか、さもなくば死ぬ)と修復能力の高さが強調でき、同時に、直接沢山の生物のゲノム解析を行うことでゲノム変異の種類や程度を調べることができる時代になったことが伝えられる点で意義深いと考えている。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (22 results)

All 2022 2021 2020 2019 2018

All Journal Article (14 results) (of which Int'l Joint Research: 5 results,  Peer Reviewed: 14 results,  Open Access: 9 results) Presentation (8 results)

  • [Journal Article] Mutations in the stator protein PomA affect switching of rotational direction in bacterial flagellar motor2022

    • Author(s)
      Terashima Hiroyuki、Hori Kiyoshiro、Ihara Kunio、Homma Michio、Kojima Seiji
    • Journal Title

      Scientific Reports

      Volume: 12 Issue: 1 Pages: 2979-2979

    • DOI

      10.1038/s41598-022-06947-5

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Two substates in the O intermediate of the light-driven proton pump archaerhodopsin-22022

    • Author(s)
      Kouyama Tsutomu、Ihara Kunio
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - Biomembranes

      Volume: 1864 Issue: 7 Pages: 183919-183919

    • DOI

      10.1016/j.bbamem.2022.183919

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification of <i>sur2</i> mutation affecting the lifespan of fission yeast2021

    • Author(s)
      Kurauchi Tatsuhiro、Matsui Kotaro、Shimasaki Takafumi、Ohtsuka Hokuto、Tsubouchi Satoshi、Ihara Kunio、Tani Motohiro、Aiba Hirofumi
    • Journal Title

      FEMS Microbiology Letters

      Volume: 368 Issue: 12

    • DOI

      10.1093/femsle/fnab070

    • NAID

      120007183219

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification of <i>ksg1</i> mutation showing long‐lived phenotype in fission yeast2021

    • Author(s)
      Matsui Kotaro、Okamoto Keisuke、Hasegawa Tomoka、Ohtsuka Hokuto、Shimasaki Takafumi、Ihara Kunio、Goto Yuhei、Aoki Kazuhiro、Aiba Hirofumi
    • Journal Title

      Genes to Cells

      Volume: 26 Issue: 12 Pages: 967-978

    • DOI

      10.1111/gtc.12897

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A slight bending of an α-helix in FliM creates a counterclockwise-locked structure of the flagellar motor in <i>Vibrio</i>2021

    • Author(s)
      Takekawa Norihiro、Nishikino Tatsuro、Yamashita Toshiki、Hori Kiyoshiro、Onoue Yasuhiro、Ihara Kunio、Kojima Seiji、Homma Michio、Imada Katsumi
    • Journal Title

      The Journal of Biochemistry

      Volume: 170 Issue: 4 Pages: 531-538

    • DOI

      10.1093/jb/mvab074

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Loss of cell wall integrity genes cpxA and mrcB causes flocculation in Escherichia coli2021

    • Author(s)
      Sugawara Keita、Toyoda Hayato、Kimura Mami、Hayasaka Shunsuke、Saito Hiromi、Kobayashi Hiroshi、Ihara Kunio、Ida Tomoaki、Akaike Takaaki、Ando Eiji、Hyodo Mamoru、Hayakawa Yoshihiro、Hamamoto Shin、Uozumi Nobuyuki
    • Journal Title

      Biochemical Journal

      Volume: 478 Issue: 1 Pages: 41-59

    • DOI

      10.1042/bcj20200723

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Presynaptic MAST kinase controls opposing postsynaptic responses to convey stimulus valence in Caenorhabditis elegans2020

    • Author(s)
      Nakano Shunji、Ikeda Muneki、Tsukada Yuki、Fei Xianfeng、Suzuki Takamasa、Niino Yusuke、Ahluwalia Rhea、Sano Ayana、Kondo Rumi、Ihara Kunio、Miyawaki Atsushi、Hashimoto Koichi、Higashiyama Tetsuya、Mori Ikue
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 117 Issue: 3 Pages: 1638-1647

    • DOI

      10.1073/pnas.1909240117

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Energetics and kinetics of substrate analog-coupled staphylococcal nuclease folding revealed by a statistical mechanical approach.2020

    • Author(s)
      Mizukami T, Furuzawa S, Itoh SG, Segawa S, Ikura T, Ihara K, Okumura H, Roder H, Maki K
    • Journal Title

      Proceedings of the National Academy of Sciences of the United States of America

      Volume: 117 Issue: 33 Pages: 19953-19962

    • DOI

      10.1073/pnas.1914349117

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] The role of ROC75 as a daytime component of the circadian oscillator in Chlamydomonas reinhardtii2020

    • Author(s)
      Matsuo Takuya, Iida Takahiro, Ohmura Ayumi, Gururaj Malavika, Kato Daisaku, Mutoh Risa, Ihara Kunio, Ishiura Masahiro
    • Journal Title

      PLOS GENETICS

      Volume: 16 Issue: 6 Pages: e1008814-e1008814

    • DOI

      10.1371/journal.pgen.1008814

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Haloarcula mannanilytica sp. nov., a galactomannan-degrading haloarchaeon isolated from commercial salt2020

    • Author(s)
      Enomoto Shigeaki, Shimane Yasuhiro, Ihara Kunio, Kamekura Masahiro, Itoh Takashi, Ohkuma Moriya, Takahashi--Ando Naoko, Fukushima Yasumasa, Yoshida Yasuhiko, Usami Ron, Takai Ken, Minegishi Hiroaki
    • Journal Title

      INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY

      Volume: 70 Issue: 12 Pages: 6331-6337

    • DOI

      10.1099/ijsem.0.004535

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] The mechanosensitive channel YbdG from Escherichia coli has a role in adaptation to osmotic up-shock.2019

    • Author(s)
      Shun Amemiya, Hayato Toyoda, Mami Kimura, Hiromi Saito, Hiroshi Kobayashi, Kunio Ihara 3 , Kiyoto Kamagata 4 , Ryuji Kawabata 5 , Setsu Kato, Yutaka Nakashimada, Tadaomi Furuta, Shin Hamamoto, Nobuyuki Uozumi
    • Journal Title

      Journal of Biological Chemistry

      Volume: 294 (33) Issue: 33 Pages: 12281-12292

    • DOI

      10.1074/jbc.ra118.007340

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Cholecystokinin induces crowing in chickens2019

    • Author(s)
      Shimmura T, Tamura M, Ohashi S, Sasaki A, Yamanaka T, Nakao N, Ihara K, Okamura S, Yoshimura T.
    • Journal Title

      Sci Rep.

      Volume: 9 Issue: 1 Pages: 3978-3978

    • DOI

      10.1038/s41598-019-40746-9

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] gas1 mutation extends chronological lifespan via Pmk1 and Sty1 MAPKs in Schizosaccharomyces pombe2019

    • Author(s)
      Imai Yuki、Shimasaki Takafumi、Enokimura Chihiro、Ohtsuka Hokuto、Tsubouchi Satoshi、Ihara Kunio、Aiba Hirofumi
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 84 Issue: 2 Pages: 330-337

    • DOI

      10.1080/09168451.2019.1676695

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Obesity and mental health improvement following nutritional education focusing on gut microbiota composition in Japanese women: a randomised controlled trial2018

    • Author(s)
      Uemura Mayu、Hayashi Fumikazu、Ishioka Ken、Ihara Kunio、Yasuda Kazushi、Okazaki Kanako、Omata Junichi、Suzutani Tatsuo、Hirakawa Yoshihisa、Chiang Chifa、Aoyama Atsuko、Ohira Tetsuya
    • Journal Title

      European Journal of Nutrition

      Volume: 2018 Issue: 8 Pages: 1-12

    • DOI

      10.1007/s00394-018-1873-0

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Presentation] 大量ゲノム解析に基づいた UVC 光照射法による変異導入効率の評価2022

    • Author(s)
      松尾佳祐、上坂一馬、井原邦夫
    • Organizer
      第16回日本ゲノム微生物学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] シアノバクテリア Leptolyngbya boryana の光合成依存的な窒素固定生育に関わる遺伝子群の探索2022

    • Author(s)
      馬場真里、上坂一馬、戸松 千映、山本 治樹 、井原 邦夫 、藤田祐一
    • Organizer
      第16回日本ゲノム微生物学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 上坂一馬, 馬場真里、 藤田祐一、 井原 邦夫2022

    • Author(s)
      Tn 挿入ライブラリからゲノム変異を網羅的に検出するワークフローの開発
    • Organizer
      第16回日本ゲノム微生物学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] UVc照射によるゲノム変異導入は効果的な方法か?2021

    • Author(s)
      井原邦夫, 松尾佳祐、上坂一馬
    • Organizer
      第59回日本生物物理学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] バクテリオロドプシンの光反応サイクルにおける2種類のO中間体の存在2021

    • Author(s)
      神山勉、井原邦夫
    • Organizer
      第59回日本生物物理学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 高度好塩性好アルカリ性古細菌 Natronomonas pharaonisを用いた大量変異導入と全ゲノムシーケンス解析の実験系の開発2021

    • Author(s)
      松尾佳祐、上坂一馬、井原邦夫
    • Organizer
      第33回日本Archaea 研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ゲノムから見直す微生物分類の再考ーーーVibrio alginolyticusの場合ーーー2021

    • Author(s)
      上坂一馬,井原邦夫
    • Organizer
      第15回 微生物ゲノム学会
    • Related Report
      2020 Research-status Report
  • [Presentation] 微生物ゲノムライブラリーを手軽で格安に作る -1コインゲノム解析を目指して2020

    • Author(s)
      井原邦夫、上坂一馬
    • Organizer
      第14回 微生物ゲノム学会
    • Related Report
      2019 Research-status Report

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Published: 2019-04-18   Modified: 2023-01-30  

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