Legionella: Methods and Protocols

Part I: Ecology, Physiology and Genetics of Legionella

1. Analysis of Iron Requirements and Siderophore Production

Denise M. Burnside, and Nicholas P Cianciotto

2. The Pathometabolism of Legionella Studied by Isotopologue Profiling

Klaus Heuner, Mareike Kunze, Fan Chen, and Wolfgang Eisenreich

3. Quantification of Viable but Non-culturable Cells of Legionella pneumophila

Rosalie S. Braun, Nilmini Mendis, Laam Li, and Sebastien P. Faucher

4. Selection of Legionella Virulence-related Traits by Environmental Protozoa

Francisco Amaro and Howard Shuman

5. Migration of Acanthamoeba castellanii through Legionella Biofilms

Ramon Hochstrasser and Hubert Hilbi

6. Scar-free Genome Editing in Legionella pneumophila

Nathalie Bailo, Hussein Kanaan, Elisabeth Kay, Xavier Charpentier, Patricia Doublet, and Christophe Gilbert

7. Querying Legionella Genomes using Transposition-sequencing

L?o Hardy and Xavier Charpentier

8. Screening Targeted Legionella pneumophila Mutant Libraries in vivo using INSeq

Stephanie R. Shames

9. Genetic Manipulation of non-pneumophila Legionella: Protocols Developed for Legionella longbeachae

Elizabeth Korevaar, Chen Ai Khoo, and Hayley J. Newton

Part II: Cellular Microbiology of Legionella

10. Quantitative Imaging Flow Cytometry of Legionella-containing Vacuoles in Dually Fluorescence-labelled Dictyostelium

Amanda Welin, Stephen Weber, and Hubert Hilbi

11. Sorting of Phagocytic Cells Infected with Legionella pneumophila

Monica Rolando and Carmen Buchrieser

12. Single Cell Analysis of Legionella and Legionella-infected Acanthamoeba by Agarose Embedment

Nicolas Personnic, Bianca Striednig, and Hubert Hilbi

13. Mitochondrial Dynamics and Activity in Legionella-Infected Cells

Ok-Ryul Song, Priscille Brodin, Carmen Buchrieser, and Pedro Escoll

14. Perturbation of Legionella Cell Infection by RNA Interference

Bernhard Steiner, A. Leoni Swart, and Hubert Hilbi

Part III: Biochemical Assays to Study Legionella Effectors and Enzymes

15. Isolation of the Dot/Icm Type IV Secretion System Core Complex from Legionella pneumophila

Tomoko Kubori and Hiroki Nagai

16. In situ Imaging and Structure Determination of Bacterial Toxin Delivery Systems using Electron Cryotomography

Debnath Ghosal, Mohammed Kaplan, Yi-Wei Chang, and Grant J. Jensen

17. Methods for Non-canonical Ubiquitination and Deubiquitination Catalyzed by Legionella pneumophila Effector Proteins

Jiazhang Qiu and Zhao-Qing Luo

18. Purification and Analysis of Effector Glucosyltransferase Lgt1 from Legionella pneumophila

Nadezhda Levanova, Irina Tabakova, Thomas Jank, and Yury Belyi

19. Determination of in vivo Interactomes of Dot/Icm type IV Secretion System Effectors by Tandem Affinity Purification

Ernest C. So, Aur?lie Mousnier, Gad Frankel, and Gunnar N. Schroeder

20. Inflammasome Activation in Legionella-infected Macrophages

Danielle P. A. Mascarenhas and Dario S. Zamboni

Part IV. Immunity and Host Response Against Legionella

21. Infection of Human Lung Tissue Ex