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Establishment of method for injection of heterologous genomic DNA into bacterial giant cells

Research Project

Project/Area Number 16K14891
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Applied microbiology
Research InstitutionToyama Prefectural University

Principal Investigator

Nishida Hiromi  富山県立大学, 工学部, 教授 (60301115)

Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsスフェロプラスト / 巨大化細胞 / マイクロマニュピレーション操作 / 金属イオン / Lelliottia amnigena / 光合成細菌 / Deinococcus grandis / マイクロインジェクション / バクテリア / 巨大化 / マイクロマニュピレータ / Rhodospirillum / Erythrobacter / Deinococcus / Lelliottia / マイクロマニピュレーション操作 / 内膜 / 外膜 / DNA複製 / 長鎖DNA導入 / マリン培地 / 細胞巨大化・脱巨大化 / 異種ゲノムDNA導入 / 細胞分裂
Outline of Final Research Achievements

Injection of foreign DNA into bacterial cells has limitation in size and it is impossible to inject chromosome size DNA (> 1 Mbp). Even if genome of bacteria is designed and DNA can be synthesized based on its design drawing, injection of the DNA into the cell is impossible. So, we aimed to establish a method to inject foreign long DNA by piercing the microchip into the cell. In order to achieve the objective, it was necessary to enlarge the bacterial cells of 1-2 μm up to >15 μm. We succeeded in enlarging several bacterial cells so far and carried out micromanipulation operation. Particularly in the giant cells of Lelliottia amnigena, noteworthy results could be raised.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (9 results)

All 2018 2017 2016

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (7 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results)

  • [Journal Article] Comparison of gene expression levels of appA, ppsR, and EL368 in Erythrobacter litoralis spheroplasts under aerobic and anaerobic conditions, and under blue light, red light, and dark conditions2018

    • Author(s)
      Nishino K, Takahashi S, Nishida H
    • Journal Title

      Journal of General and Applied Microbiology

      Volume: 64

    • NAID

      130007417999

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Effects of light and oxygen on the enlargement of spheroplasts of the facultative anaerobic anoxygenic photosynthetic bacterium Rhodospirillum rubrum2017

    • Author(s)
      Nakazawa M, Nishida H
    • Journal Title

      Jacobs Journal of Biotechnology and Bioengineering

      Volume: 3 Pages: 014-014

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] バクテリアのスフェロプラスト巨大化2018

    • Author(s)
      西田洋巳
    • Organizer
      日本農芸化学会2018年度大会シンポジウム「バクテリア細胞膜の機能および構造研究の新展開」
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] バクテリア巨大化スフェロプラストは、線維化を経て2分裂細胞に戻るか?2017

    • Author(s)
      高橋沙和子,西田洋巳
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都
    • Year and Date
      2017-03-20
    • Related Report
      2016 Research-status Report
  • [Presentation] 好気性紅色光合成細菌スフェロプラスト巨大化における呼吸と光合成の影響2017

    • Author(s)
      西野弘起,中澤舞,西田洋巳
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都
    • Year and Date
      2017-03-20
    • Related Report
      2016 Research-status Report
  • [Presentation] Quantitative analysis of DNA and RNA in enlarged spheroplasts of the bacterium Lelliottia amnigena2017

    • Author(s)
      Sawako Takahashi, Hiromi Nishida
    • Organizer
      FEMS 2017, 7th Congress of European Microbiologists
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Deinococcusのスフェロプラストに関する研究2017

    • Author(s)
      梅村幸佑,森田裕介,西野弘起,白谷周作,鳴海一成,大島拓,西田洋巳
    • Organizer
      第10回北陸合同バイオシンポジウム
    • Related Report
      2017 Annual Research Report
  • [Presentation] 腸内細菌科のLelliottia amnigena巨大化スフェロプラストの遺伝子発現解析2016

    • Author(s)
      高橋沙和子,西田洋巳
    • Organizer
      第9回北陸合同バイオシンポジウム
    • Place of Presentation
      あわら
    • Year and Date
      2016-11-04
    • Related Report
      2016 Research-status Report
  • [Presentation] 好気性紅色光合成細菌Erythrobacter litoralisのスフェロプラスト成長における光と酸素の影響2016

    • Author(s)
      中澤舞,西田洋巳
    • Organizer
      第9回北陸合同バイオシンポジウム
    • Place of Presentation
      あわら
    • Year and Date
      2016-11-04
    • Related Report
      2016 Research-status Report

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Published: 2016-04-21   Modified: 2019-03-29  

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