Abstract
The secondary messenger molecule, 3′,5′-cyclic diguanosine monophosphate (c-di-GMP), controls various cellular processes in bacteria. Direct measurement of intracellular concentration of c-di-GMP is fast becoming an important tool for studying prokaryotic biology. Here, we describe a comprehensive extraction protocol from live bacteria and quantitative analysis using LC/MS/MS.
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Acknowledgments
The authors would like to thank Dale Whittington, Ross Lawrence, and the late Thomas Kalhorn for their expertise in LC/MS/MS. This work was supported in part by NIH grant MCB0822405 and AI061396-06 to MRP. YI was a UW CFF RDP Fellow.
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Irie, Y., Parsek, M.R. (2014). LC/MS/MS-Based Quantitative Assay for the Secondary Messenger Molecule, c-di-GMP. In: Filloux, A., Ramos, JL. (eds) Pseudomonas Methods and Protocols. Methods in Molecular Biology, vol 1149. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-0473-0_22
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DOI: https://doi.org/10.1007/978-1-4939-0473-0_22
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