Bacterial Virulence: Methods and Protocols

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, C幨ine 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