2019 Fiscal Year Final Research Report
Development of large DNA delivery into chloroplast using the fusion peptide
| Project/Area Number |
17K19278
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Agricultural and Environmental Biology and related fields
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| Research Institution | Takasaki University of Health and Welfare (2018-2019)
Institute of Physical and Chemical Research (2017) |
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Principal Investigator |
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| Project Period (FY) |
2017-06-30 – 2020-03-31
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| Keywords | 葉緑体 / 形質転換 / 長鎖DNA / ペプチド / タバコ |
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| Outline of Final Research Achievements |
We developed 85 kbp plasmid containing a selection marker gene and homologous sequences of tobacco chloroplast genome. This plasmid was delivered into tobacco chloroplast by peptide-based DNA carrier which has a chloroplast targeting sequence. PCR analysis revealed that integration of the plasmid in the chloroplast genome transiently. On the other hands, the recombination was not detected with the fragmented plasmid, indicating the peptide protected the plasmid and delivered it into chloroplasts. Antibiotic-resistant tobacco shoots were regenerated from cotyledons treated with the plasmid and the peptide. Finally, 3 T0 transgenic tobaccos were grown on soil.
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| Free Research Field |
合成生物学
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| Academic Significance and Societal Importance of the Research Achievements |
近年、長鎖DNAの合成や対象生物へ導入する研究が盛んに行われている。長鎖DNAは物理的せん断に対して感受性が高く、一般的な実験で容易に断片化する。葉緑体は物質生産という観点から非常に魅力的な反応場と考えられている。しかし、一般的に葉緑体の組換えに利用されるパーティクルガン法は、物理的せん断を容易に引き起こすことが考えられるため、長鎖DNAを用いた研究は報告されていない。本研究成果は長鎖DNAを保護しつつ葉緑体を組換えることを可能とする画期的な方法となる。この方法を用いることで、複数の遺伝子を格納した長鎖DNA導入により、これまでまで生合成できなかった有用物質生産などに応用が期待できる。
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