Legionella (2nd Ed., 2nd ed. 2019)
Methods and Protocols

Methods in Molecular Biology Series, Vol. 1921

Coordinators: Buchrieser Carmen, Hilbi Hubert

Language: Anglais

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484 p. · 17.8x25.4 cm · Hardback
The methods described in the second edition on Legionella are for the study of distinct features of L. pneumophila.  Chapters guide readers through ecology and physiology of legionella, legionella genetics, cellular microbiology of legionella, biochemical assays to study legionella effectors and enzymes, immunity and host response against legionella, metagenomics, proteomics, and host microbiomes of legionella. 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.

Authoritative and cutting-edge, Legionella: Methods and Protocols, Second Edition aims to be useful for scientists studying Legionella,  and for a broader research community interested in the ecology, physiology, pathogenesis, immunity, genetics and evolution of other bacterial pathogens.

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

Léo Hardyand 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 CellsInfected with Legionella pneumophila

Monica Rolandoand 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, Aurélie 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 Explants (HLTEs) with Legionella pneumophila

Lina Scheithauer and Michael Steinert


22. The Galleria mellonella InfectionModelfor Investigating the Molecular Mechanisms of Legionella Virulence

Gad Frankeland Gunnar N. Schroeder


23. Dictyostelium Host Response to Legionella Infection: Strategies and Assays Salvatore Bozzaro, Simona Buracco, Barbara Peracino, and Ludwig Eichinger


24. 6.1.3 The Caenorhabditis elegans model of Legionella infection

Ann Karen C. Brassinga and Costi D. Sifri


25. The Mouse as a Model for Pulmonary Legionella Infection

Garrett Z. Ng, Anastasia Solomatina, Ian R. van Driel, and

Elizabeth L. Hartland


Part V. Genomics, Transcriptomics, and Proteomics of Legionella

26. Targeted Metagenomics of Microbial Diversity in Free-living Amoebae and Water Samples

Vincent Delafont, Yoann Perrin, Didier Bouchon, Laurent Moulin, and Yann Héchard


27. Analyses of the Pulmonary Microbiome Composition of Legionella Pneumophila-Infected Patients

Ana Elena Pérez-Cobas and Carmen Buchrieser

28. Subcellular Protein Fractionation in Legionella pneumophila and Preparation of the Derived Sub-proteomes for Analysis by Mass Spectrometry

Sandra Maaß, Gina Moog, and Dörte Becher

Includes cutting-edge methods and protocols 

Provides step-by-step detail essential for reproducible results 

Contains key notes and implementation advice from the experts