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.

中文翻译：

---

基于噬菌体的生物传感器：体内分析天然T4噬菌体启动子，以增强报告酶的表达。

基于噬菌体的生物传感器由于具有足够的便携性，速度，易用性，灵敏性和较低的生产成本，在满足食品和农业工业的当前需求方面显示出了巨大的希望。尽管当前基于噬菌体的方法不能满足EPA，FDA和USDA规定的细菌检测极限，但是对噬菌体遗传学的更好理解可以显着提高其作为生物传感器的敏感性。在当前的研究中，基于T4噬菌体的检测系统的信号灵敏度通过报道酶Nanoluc荧光素酶（Nluc）的转录上调。开发了一个有效的平台，以评估报告程序T4噬菌体的启动子活性。使用15个天然T4启动子评估了T4噬菌体中Nluc上调的能力。数据表明来自所选启动子整合的报告酶信号增加了六倍。总的来说，这项工作表明，通过优化转录来微调报告酶（例如Nluc）的表达可以显着降低检测限。