Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, USA.
Methods Mol Biol. 2021;2323:153-170. doi: 10.1007/978-1-0716-1499-0_12.
Riboswitches are a class of noncoding RNAs that regulate gene expression in response to changes in intracellular metabolite concentrations. When riboswitches are placed upstream of genetic reporters, the degree of reporter activity reflects the relative abundance of the metabolite that is sensed by the riboswitch. This method describes how reporters for live cell imaging, such as yellow fluorescent protein (YFP), can be placed under genetic control by metabolite-sensing riboswitches in the bacterium Bacillus subtilis. Specifically, a protocol for generating a fluorescent YFP reporter, based on a c-di-GMP responsive riboswitch, is outlined below.
Riboswitches 是一类非编码 RNA，能够响应细胞内代谢物浓度变化来调节基因表达。当 riboswitches 被放置在遗传报告基因的上游时，报告基因的活性程度反映了 riboswitch 所感应到的代谢物的相对丰度。本方法描述了如何在细菌枯草芽孢杆菌中，通过代谢物感应 riboswitches 对活体成像报告基因（如黄色荧光蛋白，YFP）进行遗传控制。具体而言，下面概述了一种基于 c-di-GMP 反应性 riboswitch 生成荧光 YFP 报告基因的方案。
