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

• Material Type: 博士論文
• Title: 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
• Author/Editor: Tabuchi Yagui, Takeshi Ricardo
• Author Heading: Tabuchi Yagui, Takeshi Ricardo
• Publication Date: 2022-09-30
• Publication Date (W3CDTF): 2022-09-30
• Alternative Title: High-Throughput Screening of Cell-Free Riboswitches for Chemical Communication between Microdroplets
• Degree grantor/type: Okinawa Institute of Science and Technology Graduate University
• Date Granted: 2022-09-30
• Date Granted (W3CDTF): 2022-09-30
• Dissertation Number: 甲第102号
• Degree Type: Doctor of Philosophy
• Conferring No. (Dissertation): 甲第102号
• Text Language Code: eng
• Target Audience: 一般
• Note (General): 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.
• DOI: 10.15102/1394.00002535; https://doi.org/10.15102/1394.00002535
• Persistent ID (NDL): info:ndljp/pid/12361287; https://dl.ndl.go.jp/pid/12361287
• Collection: 障害者向け資料
• Collection (Materials For Handicapped People:1): テキストデータ
• Collection (particular): 国立国会図書館デジタルコレクション > デジタル化資料 > 博士論文; https://dl.ndl.go.jp/collections/A00014
• Acquisition Basis: 博士論文（自動収集）
• Date Accepted (W3CDTF): 2022-11-07T16:56:35+09:00
• Date Created (W3CDTF): 2022-10-03
• Format (IMT): PDF; application/pdf
• Access Restrictions: 国立国会図書館内限定公開
• Service for the Digitized Contents Transmission Service: 図書館・個人送信対象外
• Availability of remote photoduplication service: 可
• Periodical Title (URI): http://dx.doi.org/10.15102/1394.00002535; https://oist.repo.nii.ac.jp/records/2804
• Data Provider (Database): 国立国会図書館 : 国立国会図書館デジタルコレクション; https://dl.ndl.go.jp