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RATIOMETRIC BIOSENSOR TO MEASURE INTRACELLULAR NADH/NAD+ REDOX
RATIOMETRIC BIOSENSOR TO MEASURE INTRACELLULAR NADH/NAD+ REDOX
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机译:比率生物传感器测量细胞内NADH / NAD +氧化还原
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摘要
Described herein is a redox biosensor system for measurement of NADH/NAD+ ratio including a host strain or cell-free system including first and a second expression cassette; the first expression cassette including, in operable communication, a constitutive promoter and a gene encoding a Rex allosteric transcription factor that regulates gene expression by binding to NAD(H); and the second expression cassette including, in operable communication, a promoter regulated by the Rex allosteric transcription factor, and a gene encoding a reporter protein, wherein the first expression cassette expresses the Rex allosteric transcription factor, and, in the presence of NAD(H), the Rex allosteric transcription factor drives expression of the reporter protein from the second expression cassette, and wherein the expression of the reporter protein is proportional to the NADH/NAD+ ratio.
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机译:本文描述了一种用于测量NADH / NAD + Sup>比的氧化还原生物传感器系统,其包括宿主菌株或包括第一和第二表达盒的无细胞系统。第一表达盒,包括可操作地连通的组成型启动子和编码Rex变构转录因子的基因,该基因通过与NAD(H)结合来调节基因表达;以及第二表达盒,其以可操作的方式包括受雷克斯变构转录因子调节的启动子和编码报道蛋白的基因,其中第一表达盒在不存在NAD(H)的情况下表达雷克斯变构转录因子。 ),Rex变构转录因子驱动第二表达盒中报告蛋白的表达,其中报告蛋白的表达与NADH / NAD + Sup>的比例成比例。
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