https://scholars.lib.ntu.edu.tw/handle/123456789/634285
標題: | Harnessing split fluorescent proteins in modular protein logic for advanced whole-cell detection | 作者: | Lin, Ping Heng Tsai, Ssu Tzu Chang, Yu Chia YI-JU CHOU Yeh, Yi Chun |
關鍵字: | AND gate | Heavy metal | HrpRS | Split fluorescent proteins | Spy Tag/Spy Catcher | Whole-cell biosensors | 公開日期: | 22-九月-2023 | 卷: | 1275 | 來源出版物: | Analytica Chimica Acta | 摘要: | Whole-cell biosensors have demonstrated promising capabilities in detecting target molecules. However, their limited selectivity and precision can be attributed to the broad substrate tolerance of natural proteins. In this study, we aim to enhance the performance of whole-cell biosensors by incorporating of logic AND gates. Specifically, we utilize the HrpR/S system, a widely employed hetero-regulation module from Pseudomonas syringae in synthetic biology, to construct an orthogonal AND gate in Escherichia coli. To accomplish this, we compare the HrpR/S system with self-associating split fluorescent proteins using the Spy Tag/Spy Catcher system. Our objective is to selectively activate a reporter gene in the presence of both IPTG and Hg(II) ions. Through systematic genetic engineering and evaluation of various biological parts under diverse working conditions, our research demonstrates the utility of self-associating split fluorescent proteins in developing high-performance whole-cell biosensors. This approach offers advantages such as engineering simplicity, reduced basal activity, and improved selectivity. Furthermore, the comparison with the HrpR/S system serves as a valuable control model, providing insights into the relative advantages and limitations of each approach. These findings present a systematic and adaptable strategy to overcome the substrate tolerance challenge faced by whole-cell biosensors. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/634285 | ISSN: | 00032670 | DOI: | 10.1016/j.aca.2023.341593 |
顯示於: | 應用力學研究所 |
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