期刊
ANALYTICAL CHEMISTRY
卷 91, 期 23, 页码 15284-15292出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.9b04444
关键词
-
资金
- UK BBSRC [BB/N007212/1]
- Leverhulme Trust [RPG-2015-445]
- UKRI Future Leaders Fellowship [MR/S018875/1]
- Wellcome Trust Seed Award in Science [202078/Z/16/Z]
- EPSRC/BBSRC Global Challenges Research Fund
- NATO Science for Peace and Security Programme [985042]
- Wellcome Trust [202078/Z/16/Z] Funding Source: Wellcome Trust
- BBSRC [BB/N007212/1] Funding Source: UKRI
- UKRI [MR/S018875/1] Funding Source: UKRI
Whole-cell and cell-free transcription-translation biosensors have recently become favorable alternatives to conventional detection methods, as they are cost-effective, environmental friendly, and easy to use. Importantly, the biological responses from the biosensors need to be converted into a physicochemical signal for easy detection, and a variety of genetic reporters have been employed for this purpose. Reporter gene selection is vital to a sensor performance and application success. However, it was largely based on trial and error with very few systematic side-by-side investigations reported. To address this bottleneck, here we compared eight reporters from three reporter categories, i.e., fluorescent (gfpmut3, deGFP, mCherry, mScarlet-I), colorimetric (lacZ), and bioluminescent (luxCDABE from Aliivibrio fischeri and Photorhabdus luminescens, NanoLuc) reporters, under the control of two representative biosensors for mercury- and quorum-sensing molecules. Both whole-cell and cell-free formats were investigated to assess key sensing features including limit of detection (LOD), input and output dynamic ranges, response time, and output visibility. For both whole-cell biosensors, the lowest detectable concentration of analytes and the fastest responses were achieved with NanoLuc. Notably, we developed, to date, the most sensitive whole-cell mercury biosensor using NanoLuc as reporter, with an LOD <= 50.0 fM HgCl2 30 min postinduction. For cell-free biosensors, overall, NanoLuc and deGFP led to shorter response time and lower LOD than the others. This comprehensive profile of diverse reporters in a single setting provides a new important benchmark for reporter selection, aiding the rapid development of whole-cell and cell-free biosensors for various applications in the environment and health.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据