2023 Fiscal Year Final Research Report
Proposal of a novel genotyping method for pathogenic bacteria
| Project/Area Number |
20K18923
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 58020:Hygiene and public health-related: including laboratory approach
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| Research Institution | Toyama Institute of Health |
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Principal Investigator |
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| Project Period (FY) |
2020-04-01 – 2024-03-31
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| Keywords | 遺伝子型別 / 三本鎖DNA / レジオネラ菌 |
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| Outline of Final Research Achievements |
In this study, we developed a formula for predicting polypurine/polypyrimidine (PolyR) triple-stranded DNA formation. Using this formula, it is possible to efficiently select sequences that can be genotyped and easily formed into triple-stranded DNA from PolyR sequences on the chromosomal DNA of microorganisms. Genotyping was performed using 24 different PolyR sequences on chromosomal DNA of Legionella species as a model, and the results were compared in silico with those of the conventional Sequence-based typing (SBT) method. As a result, the discrimination power (D) of this typing method (D value of 0.989) is higher resolution to that of the SBT method (D value of 0.981).
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| Free Research Field |
生物工学
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| Academic Significance and Societal Importance of the Research Achievements |
開発した三本鎖DNA形成温度を予測する式は, 実験的にも機能することが確認できた。本技術は, 選択した配列がどの様な条件下で三本鎖DNAを形成するかをPC上で容易に確認できる。このことにより, 精度の高い実験計画の立案から迅速な応用研究への展開まで幅広く活用可能であると考えられる。また, レジオネラ属菌をモデルに染色体DNA上の三本鎖DNA形成配列を解析することで, 遺伝子型別が可能であることを示した。三本鎖DNAの形成は, 熱変性を必要とせず二本鎖DNAの配列特異的に速やかに結合することから, 三本鎖DNA形成を利用する新規遺伝子型別法は, 迅速な分析法として期待できる。
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