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Institut Pasteur, Paris, France

2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens

Charlotte Keller^*1, Nora Mellouk^*1, Anne Danckaert², Roxane Simeone³, Roland Brosch³, Jost Enninga¹, Alexandre Bobard¹

¹Dynamique des Interactions Hôte Pathogène, Institut Pasteur, Paris, France, ²Imagopole, Institut Pasteur, Paris, France, ³Pathogenomique Mycobacterienne Integrée, Institut Pasteur, Paris, France

We describe a method for tracking the endomembrane rupture elicited by the intracellular bacteria Shigella flexneri and Mycobacterium tuberculosis upon host cell invasion. Our assay makes use of CCF4, a host cytoplasmic FRET probe in live or fixed cells. This reporter is degraded by an enzyme activity present on the bacterial surface.

Immunology and Infection

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus

Fabio Cafini^*1,2, Nguyen Thi Le Thuy^*3, Federico Román⁴, José Prieto⁵, Sarah Dubrac^6,7, Tarek Msadek^6,7, Kazuya Morikawa¹

¹Division of Biomedical Science, Faculty of Medicine, University of Tsukuba, ²Department of Basic Biomedical Science, Universidad Europea de Madrid, ³Human Biology Program, School of Integrative and Global Majors, University of Tsukuba, ⁴Laboratory of Nosocomial Infections, Department of Bacteriology, Centro Nacional de MicrobiologÍa, Instituto de Salud Carlos III, ⁵Division of Microbiology, Department of Medicine, School of Medicine, Universidad Complutense, ⁶Biology of Gram-Positive Pathogens, Department of Microbiology, Institut Pasteur, Paris, France, ⁷ERL3526, CNRS, Paris, France

We describe here three different protocols for the in vitro investigation of conjugation, transduction, and natural transformation in Staphylococcus aureus.