Environmental Laboratory, US Army Engineer Research and Development Center, Vicksburg, MS, United States of America.
Central Agricultural Research Center, Montana State University, Moccasin, MT, United States of America.
PLoS One. 2020 Nov 30;15(11):e0241664. doi: 10.1371/journal.pone.0241664. eCollection 2020.
RNA aptamers are relatively short nucleic acid sequences that bind targets with high affinity, and when combined with a riboswitch that initiates translation of a fluorescent reporter protein, can be used as a biosensor for chemical detection in various types of media. These processes span target binding at the molecular scale to fluorescence detection at the macroscale, which involves a number of intermediate rate-limiting physical (e.g., molecular conformation change) and biochemical changes (e.g., reaction velocity), which together complicate assay design. Here we describe a mathematical model developed to aid environmental detection of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) using the DsRed fluorescent reporter protein, but is general enough to potentially predict fluorescence from a broad range of water-soluble chemicals given the values of just a few kinetic rate constants as input. If we expose a riboswitch test population of Escherichia coli bacteria to a chemical dissolved in media, then the model predicts an empirically distinct, power-law relationship between the exposure concentration and the elapsed time of exposure. This relationship can be used to deduce an exposure time that meets or exceeds the optical threshold of a fluorescence detection device and inform new biosensor designs.
RNA 适体是相对较短的核酸序列,能与靶标高亲和力结合,与起始荧光报告蛋白翻译的核酶结合后,可作为用于在各种类型的介质中的化学检测的生物传感器。这些过程跨越了从分子尺度上的靶标结合到宏观尺度上的荧光检测,涉及到许多中间限速的物理(例如,分子构象变化)和生化变化(例如,反应速度),这些变化共同使测定设计复杂化。在这里,我们描述了一个数学模型的开发,该模型用于辅助使用 DsRed 荧光报告蛋白对六氢-1,3,5-三硝基-1,3,5-三嗪(RDX)的环境检测,但它足够通用,只要输入几个动力学速率常数的值,就有可能预测来自广泛的水溶性化学物质的荧光。如果我们将核糖开关测试种群的大肠杆菌暴露于溶解在介质中的化学物质中,那么该模型预测了暴露浓度与暴露时间之间存在经验上独特的幂律关系。该关系可用于推断出满足或超过荧光检测设备的光学阈值的暴露时间,并为新的生物传感器设计提供信息。