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  • 1
    Subject(s): Bacteria. ; Microbiology. ; Bacteria. ; Microbiology.
    In: Springer Nature eBook
    Description / Table of Contents: This book aims to provide methods, protocols, and discussion topics for those who wish to examine in depth the molecular mechanisms of adaptation and versality of bacteria and would like to envisage their evolution responses in the fast changing Antropocene.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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Pseudomonas aeruginosa: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
    Type of Medium: Online Resource
    Pages: X, 246 p. 51 illus., 40 illus. in color. , online resource.
    Edition: 1st ed. 2024.
    ISBN: 9781071634738
    Series Statement: Methods in Molecular Biology, 2721
    Language: English
    Note: CRISPR/Cas9-based genome editing of Pseudomonas aeruginosa -- Investigating Pseudomonas aeruginosa Gene Function During Pathogenesis using Mobile-CRISPRi -- Engineering green-light-responsive heterologous gene expression in Pseudomonas -- Fluorescence-based evaluation of cyclic di-GMP levels in Pseudomonas aeruginosa -- Whole-cell biosensors for qualitative and quantitative analysis of quorum sensing signal molecules and the investigation of quorum quenching agents -- A Pseudomonas aeruginosa-suitable fluorescent reporter system for analyzing small RNA-mediated regulation of target mRNAs -- The Pseudomonas aeruginosa resistome: permanent and transient antibiotic resistance, an overview -- Biosensors for inducers of transient antibiotic resistance -- Pseudomonas aeruginosa soluble pyocins as antibacterial weapons -- Assays for studying Pseudomonas aeruginosa secreted proteases -- Single microcolony diffusion analysis in Pseudomonas aeruginosa biofilms -- Broad genome sequencing of environmental and clinical strains and genotyping -- Genome-scale analysis of the structure and function of RNA pathways and networks in Pseudomonas aeruginosa -- In-depth quantitative proteomics analysis of the Pseudomonas aeruginosa secretome. Improving the predictive value of preclinical mouse models of Pseudomonas aeruginosa respiratory infection to evaluate antibiotic efficacy -- Emerging in vitro models for the study of infection and pathogenesis of Pseudomonas aeruginosa and testing of antibacterial agents.
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  • 2
    Publication Date: 2021-08-20
    Description: Pseudomonas aeruginosa is one of the most critical opportunistic pathogens in humans, able to cause both lethal acute and chronic lung infections. In previous work, we indicated that the small RNA ErsA plays a role in the regulatory network of P. aeruginosa pathogenicity in airways infection. To give further insight into the lifestyle functions that could be either directly or indirectly regulated by ErsA during infection, we reanalyzed the categories of genes whose transcription appeared dysregulated in an ersA knock-out mutant of the P. aeruginosa PAO1 reference strain. This preliminary analysis indicated ErsA as a candidate co-modulator of denitrification and in general, the anaerobiosis response, a characteristic physiologic state of P. aeruginosa during chronic infection of the lung of cystic fibrosis (CF) patients. To explain the pattern of dysregulation of the anaerobic-lifestyle genes in the lack of ErsA, we postulated that ErsA regulation could target the expression of Anr, a well-known transcription factor that modulates a broad regulon of anoxia-responsive genes, and also Dnr, required for the transcription activation of the denitrification machinery. Our results show that ErsA positively regulates Anr expression at the post-transcriptional level while no direct ErsA-mediated regulatory effect on Dnr was observed. However, Dnr is transcriptionally downregulated in the absence of ErsA and this is consistent with the well-characterized regulatory link between Anr and Dnr. Anr regulatory function is critical for P. aeruginosa anaerobic growth, both through denitrification and fermentation of arginine. Interestingly, we found that, differently from the laboratory strain PAO1, ErsA deletion strongly impairs the anaerobic growth by both denitrification and arginine fermentation of the RP73 clinical isolate, a multi-drug resistant P. aeruginosa CF-adapted strain. This suggests that P. aeruginosa adaptation to CF lung might result in a higher dependence on ErsA for the transduction of the multiple signals to the regulatory network of key functions for survivance in such a complex environment. Together, our results suggest that ErsA takes an upper place in the regulatory network of airways infection, transducing host inputs to biofilm-related factors, as underlined in our previous reports, and to functions that allow P. aeruginosa to thrive in low-oxygen conditions.
    Electronic ISSN: 1664-302X
    Topics: Biology
    Published by Frontiers Media
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