Bacterial Pathogenesis: Methods and Protocols

ISBN-13: 9781071632420 / Angielski

Pontus Nordenfelt; Mattias Collin

Opis

This detailed volume presents a diverse set of methodological approaches designed to improve our understanding of bacterial infections from a wide range of bacterial species. Beginning with biofilms and subcellular compartments, the book explores transcriptional analysis, methods for studying plasmid dynamics, and tools for phylogenetic analysis of bacterial genomes, as well as bacterial effector proteins interfering with host systems, host response analysis, and in vivo and in vitro infection models. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and up-to-date, Bacterial Pathogenesis: Methods and Protocols, Second Edition is a vital resource for researchers in the area of infection biology, as well as but not limited to, those working in the fields of microbiology, immunology, structural biology, molecular biology, genetics, imaging, and computational study.

Spis treści

Part I: Biofilms and Subcellular Compartments

 

1. Measuring Niche-Associated Metabolic Activity in Planktonic and Biofilm Bacteria

            Supradipta De and Anders P. Hakansson

 

2. Formation and Analysis of Mono-Species and Polymicrobial Oral Biofilms in Flow-Cell Models

            Jessica Neilands, Gunnel Svensäter, Gabriella Boisen, Carolina Robertsson, Claes Wickström, and Julia R. Davies

 

3. Isolation and Purification of Mycobacterial Extracellular Vesicles (EVs)

            Komal Umashankar Rao and Gabriela Godaly

 

4. Strategies to Isolate Extracellular Vesicles from Gram - and Gram + Bacteria

            Ana Rita Narciso and Marie-Stéphanie Aschtgen

 

Part II: Bacterial Genetics, Genomics, and Phylogenetics

 

5. Total Bacterial RNA Isolation and Northern Blotting Analysis

            Jens Karlsson, Hannes Eichner, and Edmund Loh

 

6. Phylogenetic Analysis of Bacterial Pathogen Genomes

            Xavier Didelot

 

7. Determination of Growth Rate and Virulence Plasmid Copy Number during Yersinia pseudotuberculosis Infection Using Droplet Digital PCR

            Tifaine Hechard and Helen Wang

 

Part III: Identification and Characterization of Bacterial Effector Proteins

 

8. Methods to Analyze the Contribution of Complement Evasion Factor (CEF) to Streptococcus pyogenes Virulence

            Haniyeh Aghababa, Jacelyn M.S. Loh, and Thomas Proft

 

9. Expression of the Bacterial Enzyme IdeS Using a GFP Fusion in the Yeast Saccharomyces cerevisiae

            Tova Lindh, Mattias Collin, Rolf Lood, and Magnus Carlquist

 

10. Mass Spectrometry-Based Methods to Determine the Substrate Specificities and Kinetics of N-Linked Glycan Hydrolysis by Endo-β-N-Acetylglucosaminidases

            Jonathan J. Du, Diego Sastre, Beatriz Trastoy, Blaine Roberts, Daniel Deredge, Erik H. Klontz, Maria W. Flowers, Nazneen Sultana, Marcelo E. Guerin, and Eric J. Sundberg

 

11. Identification of Substrates of Secreted Bacterial Protease by APEX2-Based Proximity Labeling

            Yanxuan Xie, Yang Mao, Zong-Wan Mao, and Wei Xia

 

12. Affinity-Purification Combined with Crosslinking Mass Spectrometry for Identification and Structural Modeling of Host-Pathogen Protein-Protein Complexes

            Lotta J. Happonen

 

13. Elucidating the Stoichiometries of Host-Pathogen Protein Interactions with Mass Photometry

            Emma-Jayne Proctor, Sandeep Satapathy, and Martina Sanderson-Smith

 

Part IV: Analysis of Host Responses to Bacteria

 

14. Analysis of Neutrophil and Monocyte Inflammation Markers in Response to Gram-Positive Anaerobic Cocci

            Tobias Schmidt and Ariane Neumann

 

15. Quantification of Phagocytosis Using Flow Cytometry

            Therese de Neergaard and Pontus Nordenfelt

 

16. Anti-Bacterial Neutrophil Effector Response: Ex Vivo Quantification of Regulated Cell Death Associated with Extracellular Trap Release

            Tiziano A. Schweizer, Sanne Hertegonne, Clément Vulin, Annelies S. Zinkernagel, and Srikanth Mairpady Shambat

 

17. Measurement of Antibody Binding Affinity on Bacterial Surfaces Using Flow Cytometry

            Vibha Kumra Ahnlide and Pontus Nordenfelt

 

18. Detection of Inflammasome Activation in Murine Bone Marrow-Derived Macrophages Infected with Group A Streptococcus

            Christine Valfridsson, Elsa Westerlund, Dóra Hancz, and Jenny J. Persson

 

Part V: In Vivo and In Vitro Infection Models

 

19. In Vivo Profiling of the Vascular Cell Surface Proteome in Murine Models of Bacteremia

            Charlotte Spliid, Jeffrey D. Esko, Johan Malmström, and Alejandro Gomez Toledo

 

20. Barcoded Consortium Infections: A Scalable, Internally Controlled Method to Study Host Cell Binding and Invasion by Pathogenic Bacteria

            Maria Letizia Di Martino and Mikael E. Sellin

 

21. A Murine Mycobacterium marinum Infection Model for Longitudinal Analyses of Disease Development and the Inflammatory Response

            Julia Lienard, Kristina Munke, and Fredric Carlsson

 

22. In Vitro Approaches for the Study of Pneumococcal-Neuronal Interaction and Pathogenesis

            Kristine Farmen and Miguel Tofiño-Vian