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Development of high sencitive FISH for revealing functions of uncultured microorganisms

Research Project

Project/Area Number 15K14807
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Aquatic life science
Research InstitutionAnan National College of Technology

Principal Investigator

Kawakami Shuji  阿南工業高等専門学校, 創造技術工学科, 講師 (00610461)

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: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords環境微生物 / シングルセル解析 / FISH法 / aptamer / 微生物 / 排水処理 / アプタマー / 未培養微生物
Outline of Final Research Achievements

In this study, we introduced an improved in situ DNA-HCR protocol (quickHCR-FISH) with higher sensitivity, and the amplification time was accelerated to only 15 min by using a dextran sulfate and blocking reagent containing amplification buffer, compared with 2 h in the original protocol. Finally, quickHCR-FISH was applied to marine bacteria in seawater and sediment samples for checking applicability of environmental microorganisms. The detection rates of marine bacteria after quickHCR-FISH were almost the same as after CARD-FISH. This newly developed protocol can be an attractive alternative to CARD-FISH for the detection of microorganisms in environmental context.

Report

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

    (4 results)

All 2017 2016 2015

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Acknowledgement Compliant: 1 results) Presentation (3 results)

  • [Journal Article] Rapid and sensitive identification of marine bacteria by an improved in situ DNA hybridization chain reaction (quickHCR-FISH)2015

    • Author(s)
      Tsuyoshi Yamaguchi, Bernhard Maximilian Fuchs, Rudolf Amann, Shuji Kawakami, Kengo Kubota, Masashi Hatamoto and Takashi Yamaguchi.
    • Journal Title

      Systematic and Applied Microbiology

      Volume: 38 Pages: 400-405

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] 蛍光標識 aptamer による E._coli の視覚的検出~高感度化と蛍光の均一化の検討~2017

    • Author(s)
      川上周司, 山口剛士, 山田剛史, 山口隆司
    • Organizer
      第51回水環境学会年会
    • Place of Presentation
      熊本大学
    • Year and Date
      2017-03-15
    • Related Report
      2016 Annual Research Report
  • [Presentation] Click_chemistry を利用した新規高感度 FISH 法の開発2017

    • Author(s)
      木村圭佑, 山口剛士, 山口隆司, 川上周司
    • Organizer
      第51回水環境学会年会
    • Place of Presentation
      熊本大学
    • Year and Date
      2017-03-15
    • Related Report
      2016 Annual Research Report
  • [Presentation] Click chemistry を用いた新規高感度 FISH 法の開発2016

    • Author(s)
      木村圭佑,山口剛士,山口隆司,川上周司
    • Organizer
      第50回日本水環境学会年会
    • Place of Presentation
      アスティ徳島
    • Year and Date
      2016-03-16
    • Related Report
      2015 Research-status Report

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

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