Part I: Molecular Biology and Bioinformatics Methods
1. Generation of Markerless Gene Deletion Mutants in Listeria monocytogenes Using a Mutated PheS for Counterselection
Shai Ran Sapir, Etai Boichis, and Anat A. Herskovits
2. A Rapid Fluorescence-Based Screen to Identify Regulators and Components of Interbacterial Competition Mechanisms in Bacteria
Daniel Tchelet and Dor Salomon
3. Predicting Type III Effector Proteins Using the Effectidor Web-Server
Naama Wagner, Doron Teper, and Tal Pupko
4. Assay for Type III Secretion in Escherichia coli
Bosko Mitrovic and Neta Sal-Man
5. Profiling of Secreted Type 3 Secretion System Substrates by Salmonella enterica
Rivka Shem-Tov and Ohad Gal-Mor
Part II: Cell Culture and Organoid Models of Infection
6. Analysis of SPI-1 Dependent Type III Secretion and Injection Using a NanoLuc Luciferase-Based Assay
Sara Vilela Pais, Sibel Westerhausen, Erwin Bohn, and Samuel Wagner
7. Mycobacterium tuberculosis Infection of THP-1 Cells: A Model for High Content Analysis of Intracellular Growth and Drug Susceptibility
Leah Rankine-Wilson, Céline Rens, Henok Asfaw Sahile, and Yossef Av-Gay
8. Bone Marrow-Derived Macrophage (BMDM) Infection by Listeria monocytogenes
Etai Boichis, Shai Ran Sapir, and Anat A. Herskovits
9. Preparation and Inflammasome Activation of Murine Bone Marrow-Derived and Resident Peritoneal Macrophages
Izumi Sasaki and Tsuneyasu Kaisho
10. Flow Cytometry-Based Single Cell Analyses of Bacterial Adaption to Intracellular Environments
Marc Schulte and Michael Hensel
11. Quantification of Microbial Fluorescent Sensors during Live Intracellular Infections
Erez Mills and Erik Petersen
12. Dissecting Human Blood Immune Cells Response to Intracellular Infection Using Single Cell RNA-Sequencing
Shelly Hen-Avivi and Roi Avraham
13. Salmonella enterica Infection of Human and Mouse Colon Organoid-Derived Monolayers
Erin C. Boyle, Eva J. Wunschel, and Guntram A. Grassl
Part III: In Vivo Models of Infection
14. Analysis of Salmonella enterica Adhesion to Leaves of Corn Salad or Lettuce
Laura Elpers and Michael Hensel
15. Methods for Using the Galleria mellonella Invertebrate Model to Probe Enterococcus faecalis Pathogenicity
Ling Ning Lam, Debra N. Brunson, Jessica K. Kajfasz, an
This detailed volume explores the field of bacterial virulence and the effort to understand how microbial interaction with a host results in the pathology of a specific disease. This collection of selected protocols includes advanced molecular biology and bioinformatics methods, cell culture and organoid models of infection, as well as in vivo infection models that are useful to study the interaction of pathogens with plants, insects, avian, and mammalian hosts. Written for the highly successful <i>Methods in Molecular Biology </i>series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. <div><br></div><div>Authoritative and beneficial, <i>Bacterial Virulence: Methods and Protocols</i> serves as an ideal guide for researchers seeking to promote and further develop the exciting and continuously evolving field of bacterial virulence.</div><div><br></div><div>Chapter 19 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.<br></div>