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.
Preface...
Table of Contents...
Contributing Authors...
1. Measurement of Staphylococcus aureus Pigmentation by Methanol Extraction
Leia E. Sullivan and Kelly C. Rice
2. Analysis of Murein Hydrolases and Proteases through Zymography
Nichole A. Seawell and Jeffrey L. Bose
3. Detection and Quantification of Secreted Nuclease Activity in Staphylococcus aureus Culture Supernatants
Richard E. Wiemels, Rebecca A. Keogh, and Ronan K. Carroll
4. Quantitative Hemolysis Assays
Miranda J. Ridder, Seth M. Daly, Pamela R. Hall, and Jeffrey L. Bose
5. In Vitro Assay for Quantifying Clumping of Staphylococcus aureus
Heidi A. Crosby, Jakub M. Kwiecinski, and Alexander R. Horswill
6. Measuring Staphylococcal Promoter Activities using a Codon-Optimized ß-galactosidase Reporter
Christina N. Krute, Nichole A. Seawell, and Jeffrey L. Bose
7. Evaluation of Staphylococcus aureus Antibiotic Tolerance using Kill Curve Assays
Jessica N. Brandwein and Kelly C. Rice
8. Fluorescence Polarization (FP) Assay for Measuring Staphylococcus aureus Membrane Fluidity
Kiran B. Tiwari, Suranjana Sen, Craig Gatto, and Brian J. Wilkinson
9. Quantification of Staphylococcus aureus Biofilm Formation by Crystal
Violet and Confocal Microscopy
Adam B. Grossman, Dylan J. Burgin, and Kelly C. Rice
10. Growth of Staphylococcus aureus using a Rotary Cell Culture System
Matthew R. Hauserman and Kelly C. Rice
11. Time-Resolved Fluorescence Assay for Measuring Oxygen Consumption Rates in Staphylococcus aureus
Logan L. Bulock, Jennifer L. Endres, and Marat R. Sadykov
12. Quantifying Staphylococcus aureus Membrane Potential using Flow Cytometry
Neal D. Hammer
13. Best Practices for Preparation of Staphylococcus aureus Metabolomics Samples
Kimberly L. James and Kelly C. Rice
14. A Rat Model of Orthopedic Implant-Associated Infection for Identification of Staphylococcal Biofilm Proteins
Mei G. Lei, Ravi Kr. Gupta, and Chia Y. Lee
15. An Ex Vivo Model for Assessing Growth and Survivability of Staphylococcus aureus in Whole Human Blood
Brittney D. Gimza, Stephanie M. Marroquin, and Lindsey N. Shaw
16. Bone Marrow-Derived Macrophage Infection Assay
Miranda J. Ridder, Mary A. Markiewicz, and Jeffrey L. Bose
17. Measurement of Osteoblast Cytotoxicity Induced by S. aureus Secreted Toxins
Caleb Ford and James E. Cassat
18. Human Skin In Vitro Colonization Model for a Skin Wound Infected by Staphylococcus aureus Biofilm
Jenelle Chapman and Michael E. Olson
Subject Index List...
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