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This volume details the phenotypic characterization of Staphylococcus aureus, with a focus on in vitro and ex vivo methodologies. The chapters in this book cover topics such as in vitro assessment of classical S. aureus virulence attributes; quantifying promoter activity using a S. aureus codon-optimized lacZ plasmid; biologically-relevant growth environments; metabolic and stress resistance assays; and in vivo and ex vivo models of host-pathogen interaction. 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and authoritative, Staphylococcus aureus: Methods and Protocols is a valuable resource for anyone interested in this fascinating and developing field.
List of contents
Measurement of Staphylococcus aureus Pigmentation by Methanol Extraction.- Analysis of Murein Hydrolases and Proteases through Zymography.- Detection and Quantification of Secreted Nuclease Activity in Staphylococcus aureus Culture Supernatants.- Quantitative Hemolysis Assays.- In Vitro Assay for Quantifying Clumping of Staphylococcus aureus.- Measuring Staphylococcal Promoter Activities using a Codon-Optimized beta-galactosidase Reporter.- Evaluation of Staphylococcus aureus Antibiotic Tolerance using Kill Curve Assays.- Fluorescence Polarization (FP) Assay for Measuring Staphylococcus aureus Membrane Fluidity.- Quantification of Staphylococcus aureus Biofilm Formation by Crystal Violet and Confocal Microscopy.- Growth of Staphylococcus aureus using a Rotary Cell Culture System.- Time-Resolved Fluorescence Assay for Measuring Oxygen Consumption Rates in Staphylococcus aureus.- Quantifying Staphylococcus aureus Membrane Potentialusing Flow Cytometry.- Best Practices for Preparation of Staphylococcus aureus Metabolomics Samples.- A Rat Model of Orthopedic Implant-Associated Infection for Identification of Staphylococcal Biofilm Proteins.- An Ex Vivo Model for Assessing Growth and Survivability of Staphylococcus aureus in Whole Human Blood.- Bone Marrow-Derived Macrophage Infection Assay.- Measurement of Osteoblast Cytotoxicity Induced by S. aureus Secreted Toxins.- Human Skin In Vitro Colonization Model for a Skin Wound Infected by Staphylococcus aureus Biofilm.
