Project description:The near-minimal bacterium Mesoplasma florum constitutes an interesting model for synthetic genomics and systems biology studies due to its small genome, fast growth rate, and lack of pathogenic potential. However, some fundamental aspects of its biology remain largely unexplored. Here, we report a broad yet remarkably detailed characterization of M. florum by combining a wide variety of experimental approaches. More specifically, we investigated several physical and physiological aspects of this bacterium, and performed the first genome-wide analysis of its transcriptome and proteome using RNA sequencing techniques and two-dimensional liquid chromatography-tandem mass spectrometry. These analyses notably revealed the M. florum consensus promoter, a first experimental cartography of transcription units, as well as the transcription and expression levels of all annotated genes. We also converted gene transcription and expression levels into absolute molecular abundances using biomass quantification results, generating an unprecedented view of the M. florum cellular composition and functions. These characterization efforts will provide an experimental foundation for the development of a genome-scale metabolic model for M. florum and will guide future genome engineering endeavours in this simple organism.