Borlee Bradley R, Borlee Grace I, Martin Kevin H, Irie Yasuhiko
Department of Microbiology, Immunology and Pathology, Infectious Disease Research Center, Colorado State University, 200 West Lake Street, 2025 Campus Delivery, Fort Collins, CO, 80523, USA.
Department of Biology, University of Dayton, 300 College Park, Dayton, OH, 45469-2320, USA.
Methods Mol Biol. 2017;1657:99-110. doi: 10.1007/978-1-4939-7240-1_9.
3',5'-cyclic diguanosine monophosphate (cyclic di-GMP) is a bacterial secondary messenger molecule that regulates many important cellular activities and behaviors, such as motility and biofilm formation. While mass spectrometry protocols for quantitative analyses of intracellular cyclic di-GMP concentrations have been developed, they are time intensive, expensive, low-throughput, and incapable of directly monitoring dynamic changes in vivo. In this protocol, we provide a Pseudomonas aeruginosa-specific detailed methodology to assay the intracellular levels of cyclic di-GMP using biological reporters.
3',5'-环二鸟苷单磷酸（环二鸟苷酸）是一种细菌第二信使分子，可调节许多重要的细胞活动和行为，如运动性和生物膜形成。虽然已经开发出用于定量分析细胞内环二鸟苷酸浓度的质谱分析方法，但这些方法耗时、昂贵、通量低，且无法直接监测体内的动态变化。在本方法中，我们提供了一种针对铜绿假单胞菌的详细方法，用于使用生物报告基因测定细胞内环二鸟苷酸的水平。
