Phage-based biosensors: in vivo analysis of native T4 phage promoters to enhance reporter enzyme expression

Phage-based biosensors have shown significant promise in meeting the present needs of the food and agricultural industries due to a combination of sufficient portability, speed, ease of use, sensitivity, and low production cost. Although current phage-based methods do not meet the bacteria detection...

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Bibliographic Details
Published in:Analyst (London) 2020-09, Vol.145 (19), p.6291-6297
Main Authors: Duong, Michelle M, Carmody, Caitlin M, Nugen, Sam R
Format: Article
Language:English
Subjects:
Bacteriophage T4 - genetics
Biosensing Techniques
Biosensors
Enzymes
Luciferases
Optimization
Phages
Production costs
Promoter Regions, Genetic
Sensitivity
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Summary:Phage-based biosensors have shown significant promise in meeting the present needs of the food and agricultural industries due to a combination of sufficient portability, speed, ease of use, sensitivity, and low production cost. Although current phage-based methods do not meet the bacteria detection limit imposed by the EPA, FDA, and USDA, a better understanding of phage genetics can significantly increase their sensitivity as biosensors. In the current study, the signal sensitivity of a T4 phage-based detection system was improved via transcriptional upregulation of the reporter enzyme Nanoluc luciferase (Nluc). An efficient platform to evaluate the promoter activity of reporter T4 phages was developed. The ability to upregulate Nluc within T4 phages was evaluated using 15 native T4 promoters. Data indicates a six-fold increase in reporter enzyme signal from integration of the selected promoters. Collectively, this work demonstrates that fine tuning the expression of reporter enzymes such as Nluc through optimization of transcription can significantly reduce the limits of detection.
ISSN:0003-2654
1364-5528
DOI:10.1039/d0an01413c