Discovery of the Intracellular Signaling Molecule Cyclic di-GMP -- Enzymatic Production of C-di-GMP Using a Thermophilic Diguanylate Cyclase -- Synthesis of [32P]-c-di-GMP for Diguanylate Cyclase and Phosphodiesterase Activity Determinations -- High-performance Liquid Chromatography (HPLC)-based Detection and Quantitation of Cellular c-di-GMP -- Identification and Quantification of Cyclic Di-Guanosine Monophosphate and its Linear Metabolites by Reversed-Phase LC-MS/MS -- Detection of Cyclic Dinucleotides by STING -- Spectrophotometric and Mass Spectroscopic Methods for the Quantification and Kinetic Evaluation of In Vitro c-di-GMP Synthesis -- Gauging and Visualizing c-di-GMP Levels in Pseudomonas aeruginosa using Fluorescence-based Biosensors -- Cyclic di-GMP-responsive Transcriptional Reporter Bioassays in Pseudomonas aeruginosa -- Live flow cytometry analysis of c-di-GMP levels in Single Cell Populations -- Experimental Detection and Visualization of the Extracellular Matrix in Macrocolony Biofilms -- Congo Red Stain Identifies Matrix Overproduction and is an Indirect Measurement for c-di-GMP in many Species of Bacteria -- Type IV pili-dependent Motility as a Tool to Determine the Activity of c-di-GMP Modulating Enzymes in Myxococcus xanthus -- Using Light-Activated Enzymes for Modulating Intracellular c‑di-GMP Levels in Bacteria -- Analysis of c-di-GMP Levels Synthesized by a Photoreceptor Protein in Response to Different Light Qualities using an in vitro Enzymatic Assay -- Probing the Role of Cyclic di-GMP Signalling Systems in Disease using Chinese Radish -- Contribution of Cyclic di-GMP in the Control of Type III and Type VI Secretion in Pseudomonas aeruginosa -- Semi-quantitative Analysis of the Red, Dry, and Rough Colony Morphology of Salmonella enterica serovar Typhimurium and Escherichia coli using Congo Red -- Fluorescent 2-aminopurine c-di-GMP and GpG analogs as PDE Probes -- Measuring Cyclic Diguanylate (c-di-GMP)-specific Phosphodiesterase Activity using the MANT-c-di-GMP Assay -- Determining Phosphodiesterase Activity (Radioactive Assay) -- Determining Diguanylate Cyclase Activity (Radioactive Assay) -- Detection of c-di-GMP-responsive DNA Binding -- Use of Nonradiochemical DNAse Footprinting to Analyze c-di-GMP Modulation of DNA-binding Proteins -- Detection of Cyclic di-GMP Binding Proteins Utilizing a Biotinylated Cyclic di-GMP Pulldown Assay -- Probing Protein-protein Interactions with Genetically Encoded Photo-activatable Crosslinkers -- Identification of c-di-AMP-Binding Proteins Using Magnetic Beads -- Pulldown with a c-di-GMP-specific Capture Compound Coupled to Mass Spectrometry as a Powerful Tool to Identify Novel Effector Proteins -- Identification of c-di-GMP-responsive Riboswitches -- Isothermal Titration Calorimetry to Determine Apparent Dissociation Constants (Kd) and Stoichiometry of Interaction (n) of c-di-GMP Binding Proteins -- Targeting c-di-GMP Signaling, Biofilm Formation and Bacterial Motility With Small Molecules -- Discovering Selective Diguanylatecyclases Inhibitors: From PleD to Discrimination of the Active Site of cyclic-di-GMP Phosphodiesterases -- High throughput Screening for Compounds that Modulate the Cellular c-di-GMP Level in Bacteria -- Genetic Tools to Study c-di-GMP-dependent Signaling in Pseudomonas aeruginosa. .

This volume provides a collection of protocols for the common experimental approaches used in the in the burgeoning field of c-di-GMP-dependent signaling. The chapters, divided into eight major parts, guide readers through methods on synthesis, detection, quantitation, modulation of the levels of c-di-GMP present in cells, procedures to detect and evaluate the interaction of c-di-GMP, and up and coming approaches focusing on the inhibition of c-di-GMP signaling.Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.  Authoritative and cutting-edge, c-di-GMP Signaling: Methods and Protocols aims to inspire researchers to try new approaches.