Development of vaculovirus gene transfer system for fish reproductive studies

2019 Fiscal Year Final Research Report

• Project/Area Number: 16K14979
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Aquatic life science
• Research Institution: Hokkaido 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
• 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

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.

Free Research Field

魚類生殖生理学

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。