Bacterial Pathogenesis Methods and Protocols

Measuring Niche-Associated Metabolic Activity in Planktonic and Biofilm Bacteria.- Formation and Analysis of Mono-Species and Polymicrobial Oral Biofilms in Flow-Cell Models.- Isolation and Purification of Mycobacterial Extracellular Vesicles (EVs).- Strategies to Isolate Extracellular Vesicles from Gram - and Gram + Bacteria.- Total Bacterial RNA Isolation and Northern Blotting Analysis.- Phylogenetic Analysis of Bacterial Pathogen Genomes.- Determination of Growth Rate and Virulence Plasmid Copy Number during Yersinia pseudotuberculosis Infection Using Droplet Digital PCR.- Methods to Analyze the Contribution of Complement Evasion Factor (CEF) to Streptococcus pyogenes Virulence.- Expression of the Bacterial Enzyme IdeS Using a GFP Fusion in the Yeast Saccharomyces cerevisiae.- Mass Spectrometry-Based Methods to Determine the Substrate Specificities and Kinetics of N-Linked Glycan Hydrolysis by Endo-ß-N-Acetylglucosaminidases.- Identification of Substrates of Secreted Bacterial Protease by APEX2-Based Proximity Labeling.- Affinity-Purification Combined with Crosslinking Mass Spectrometry for Identification and Structural Modeling of Host-Pathogen Protein-Protein Complexes.- Elucidating the Stoichiometries of Host-Pathogen Protein Interactions with Mass Photometry.- Analysis of Neutrophil and Monocyte Inflammation Markers in Response to Gram-Positive Anaerobic Cocci.- Quantification of Phagocytosis Using Flow Cytometry.- Anti-Bacterial Neutrophil Effector Response: Ex Vivo Quantification of Regulated Cell Death Associated with Extracellular Trap Release.- Measurement of Antibody Binding Affinity on Bacterial Surfaces Using Flow Cytometry.- Detection of Inflammasome Activation in Murine Bone Marrow-Derived Macrophages Infected with Group A Streptococcus.- In Vivo Profiling of the Vascular Cell Surface Proteome in Murine Models of Bacteremia.- Barcoded Consortium Infections: A Scalable, Internally Controlled Method to Study Host Cell Binding and Invasion by Pathogenic Bacteria.- A Murine Mycobacterium marinum Infection Model for Longitudinal Analyses of Disease Development and the Inflammatory Response.- In Vitro Approaches for the Study of Pneumococcal-Neuronal Interaction and Pathogenesis.