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高スループットスクリーニングによるセルフリーリボスイッチの開発と微小液滴間化学通信への応用
https://doi.org/10.15102/1394.00002535
https://doi.org/10.15102/1394.00002535ed1e1206-2447-49c9-ba7d-61ddeaa6f3a4
| 名前 / ファイル | ライセンス | アクション |
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Yagui-FullText (9.9 MB)
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Yagui-FinalExamAbstract (42.9 kB)
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| Item type | 学位論文 / Thesis or Dissertation(1) | |||||||
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| 公開日 | 2022-10-03 | |||||||
| タイトル | ||||||||
| タイトル | 高スループットスクリーニングによるセルフリーリボスイッチの開発と微小液滴間化学通信への応用 | |||||||
| 言語 | ja | |||||||
| タイトル | ||||||||
| タイトル | High-Throughput Screening of Cell-Free Riboswitches for Chemical Communication between Microdroplets | |||||||
| 言語 | en | |||||||
| 言語 | ||||||||
| 言語 | eng | |||||||
| 資源タイプ | ||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_db06 | |||||||
| 資源タイプ | doctoral thesis | |||||||
| ID登録 | ||||||||
| ID登録 | 10.15102/1394.00002535 | |||||||
| ID登録タイプ | JaLC | |||||||
| アクセス権 | ||||||||
| アクセス権 | open access | |||||||
| アクセス権URI | http://purl.org/coar/access_right/c_abf2 | |||||||
| 著者 (英) |
Tabuchi Yagui, Takeshi Ricardo
× Tabuchi Yagui, Takeshi Ricardo
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| 抄録 | ||||||||
| 内容記述タイプ | Other | |||||||
| 内容記述 | Riboswitches have recently attracted the attention of synthetic biologists as an alternative to transcription factors for genetic regulation because of their engineerability, relative simplicity, and potential for responding to a wide array of chemical signals. However, biological constraints such as cell permeability, metabolic stability, and toxicity of their chemical ligands have prevented the development of some of those devices using conventional approaches with living cells. Cell-free systems are generally not subject to such constraints and offer a unique platform for building biochemical and genetic systems that display complex functions without using living cells. Efforts to engineer regulatory components directly in cell-free systems thus far have been based on low-throughput experimental approaches, limiting the availability of basic components for building genetically programmed cell-free systems. Here, I report a high-throughput screening method for engineering riboswitches directly in a cell-free system. Fluorescence-activated droplet-sorting (FADS) of randomized riboswitch libraries in a cell-free protein synthesis (CFPS) system rapidly identified cell-free riboswitches that respond to histamine and ciprofloxacin, compounds that are normally not compatible with conventional bacterial screening methods. Finally, the riboswitches obtained through this method were used to demonstrate chemical communication between microdroplets in a self-contained system. | |||||||
| 口頭試問日 | ||||||||
| 値 | 2022-08-05 | |||||||
| 学位授与年月日 | ||||||||
| 学位授与年月日 | 2022-09-30 | |||||||
| 学位名 | ||||||||
| 学位名 | Doctor of Philosophy | |||||||
| 学位授与番号 | ||||||||
| 学位授与番号 | 甲第102号 | |||||||
| 学位授与機関 | ||||||||
| 学位授与機関識別子Scheme | kakenhi | |||||||
| 学位授与機関識別子 | 38005 | |||||||
| 学位授与機関名 | Okinawa Institute of Science and Technology Graduate University | |||||||
| 著者版フラグ | ||||||||
| 出版タイプ | VoR | |||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||
| 権利 | ||||||||
| 権利情報 | © 2022 The Author. | |||||||
