Abstract
Biofilms are complexes structures composite by different or similar bacteria that form communities, enclosed into a polymeric extracellular matrix (EPS), constituted by polysaccharides, proteins and nucleic acids. These structures are very present on the natural environment and exert an important role on ecologic and human life. Their formation regulatory networks are composite by environment and intracellular stimuli integration, able to drastically alter the bacterial gene expression, converting their motility behavior to a sessile. In this way, to understand better this genetic conversion is extremely important. Therefore, this project aims to investigate the role of the Global Transcription Factors Fis, IHF, CRP, FNR, ArcA, NarL and Lrp on the promoters of important biofilm regulators (such as rpoS, rpoE, csgD, cpxR, flhD, fliA, yeaJ, ompR, adrA and rpoD). This will be done under different experimental conditions, as pH variation, nutrients availability, temperature changes. Besides that, we will investigate the role of the regulatory regions 5'UTR aiming to understand on an integrated way the effect of post-transcriptional mechanism on the biofilm regulatory network. For this, we will analyze the gene expression variations levels, observing the GFP production and gene expression differential profile, using RNAseq. Besides evaluate the global TFs influences on the general biofilm gene expression, we will evaluate their functions on the three biofilm composition zones, utilizing confocal fluorescence microscopy and microdissection followed by RNA sequencing for verify the gene expression differences between the three zones. (AU)
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