Protocol to identify the signaling network of nucleotide second messengers in Shigella sonnei.

Here, we present a protocol to detect the signaling network composed of cyclic AMP (cAMP) and cyclic di-GMP (c-di-GMP) signaling systems in Shigella sonnei. Technical assays include microscale thermophoresis (MST), quantitative reverse-transcription PCR (RT-qPCR), ultra-high-performance liquid chromatography-mass spectrometry (LC-MS), electrophoretic mobility shift assay (EMSA), bacterial adenylate cyclase two-hybrid (BACTH) assay, and molecular docking. For complete details on the use and execution of this protocol, please refer to Wang et al.(1).