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Development of vaculovirus gene transfer system for fish reproductive studies

Research Project

Project/Area Number 16K14979
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Aquatic life science
Research InstitutionHokkaido University

Principal Investigator

Ijiri Shigeho  北海道大学, 水産科学研究院, 准教授 (90425421)

Project Period (FY) 2016-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywordsバキュロウイルス / 遺伝子導入 / 魚類細胞 / ウナギ / 濾胞刺激ホルモン / FSH / ティラピア
Outline of Final Research Achievements

Vaculovirus gene transfer system into fish cells, that were usually found difficulty in transfection, was attempted in this study. In eel kidney derived cell line (EK1 cell), vaculovirus system enabled to introduce exogenous gene into host genome without using transposon gene transfer system. Using this novel method, we could not complete to introduce eel follicle stimulating hormone gene into EK1 genome. Further technical improvement would be necessary to establish EK1 cells that stably produce eel hormone. This system did not complete to introduce gene into live fish tissues and fish embryo, which indicates further technical development would be required. In conclusion, a method which enable to introduce exogenous gene into EK1 genome was established at the first time. This novel method is a revolutionary method which enables to construct EK1 cells that produce desired protein in a stable fashion.

Academic Significance and Societal Importance of the Research Achievements

従来法ではEK1細胞ゲノムに外来遺伝子を導入することはできなかった。バキュロウイルスによりトランスポゾン系を用いなくともEK1細胞ゲノムに外来遺伝子を高い効率で組み込むことに成功した意義は大きい。予想外の結果であり、ゲノムへの遺伝子導入に関して新たな研究展開が開かれた。
トランスポゾンを必要としないことからEK1細胞ゲノムへの外来遺伝子導入は技術的に容易となった。本研究では道半ばではあるが、ウナギ濾胞刺激ホルモン産生EK1細胞が作製されれば、新しい人為催熟法が確立される。これを例に、ウナギの生体内に様々な外来遺伝子由来のタンパク質を導入できると期待され、様々な応用研究が可能となる。

Report

(5 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (2 results)

All 2020 2017

All Presentation (1 results) Book (1 results)

  • [Presentation] バキュロウイルス遺伝子導入系を利用したGFP産生EK1細胞の作製2020

    • Author(s)
      陳顧慧・谷口祐介・伴戸久徳・笠井久会・井尻成保・足立伸次
    • Organizer
      日本水産学会春季大会
    • Related Report
      2019 Annual Research Report
  • [Book] 魚類学 第14章 生殖2017

    • Author(s)
      矢部衞、桑村哲生
    • Total Pages
      377
    • Publisher
      恒星社厚生閣
    • ISBN
      9784769916109
    • Related Report
      2017 Research-status Report

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Published: 2016-04-21   Modified: 2021-02-19  

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