Lünse Christina E, Mayer Günter
Life and Medical Sciences Institute, University of Bonn, Bonn, Germany.
Methods Mol Biol. 2014;1103:199-209. doi: 10.1007/978-1-62703-730-3_15.
Ribsowitches are putative drug targets as they often regulate the expression of essential bacterial genes. This finding necessitates the development of suitable assays, at best high-throughput (HT) compatible, which allow the screening of compound libraries for riboswitch activation. Here, we describe a HT-compatible fluorescence-based screening assay employing a minimal core motif of the Bacillus subtilis glmS riboswitch and the metabolite-induced self-cleavage assay using the full-length glmS ribozyme of Staphylococcus aureus for the identification of artificial molecules activating this regulatory RNA.
核糖开关是潜在的药物靶点,因为它们常常调控细菌必需基因的表达。这一发现使得开发合适的检测方法成为必要,最好是与高通量(HT)兼容的方法,以便能够筛选化合物文库以激活核糖开关。在此,我们描述了一种基于荧光的高通量兼容筛选检测方法,该方法采用枯草芽孢杆菌glmS核糖开关的最小核心基序,以及利用金黄色葡萄球菌全长glmS核酶的代谢物诱导自我切割检测方法,用于鉴定激活这种调控RNA的人工分子。