title: Generation and multi-phenotypic high-content screening of Coxiella burnetii transposon mutants.
abstract: Invasion and colonization of host cells by bacterial pathogens depend on the activity of a large number of prokaryotic proteins, defined as virulence factors, which can subvert and manipulate key host functions. The study of host/pathogen interactions is therefore extremely important to understand bacterial infections and develop alternative strategies to counter infectious diseases. This approach however, requires the development of new high-throughput assays for the unbiased, automated identification and characterization of bacterial virulence determinants. Here, we describe a method for the generation of a GFP-tagged mutant library by transposon mutagenesis and the development of high-content screening approaches for the simultaneous identification of multiple transposon-associated phenotypes. Our working model is the intracellular bacterial pathogen Coxiellaburnetii, the etiological agent of the zoonosis Q fever, which is associated with severe outbreaks with a consequent health and economic burden. The obligate intracellular nature of this pathogen has, until recently, severely hampered the identification of bacterial factors involved in host pathogen interactions, making of Coxiella the ideal model for the implementation of high-throughput/high-content approaches.
authors.affiliation: Cell Biology of Bacterial Infections, CNRS, FRE3689, CPBS, Université Montpellier.
authors.forename: Eric
authors.initials: E
authors.lastname: Martinez
authors.affiliation: Cell Biology of Bacterial Infections, CNRS, FRE3689, CPBS, Université Montpellier.
authors.forename: Franck
authors.initials: F
authors.lastname: Cantet
authors.affiliation: Cell Biology of Bacterial Infections, CNRS, FRE3689, CPBS, Université Montpellier; matteo.bonazzi@cpbs.cnrs.fr.
authors.forename: Matteo
authors.initials: M
authors.lastname: Bonazzi
mesh_terms: D016997:Coxiella burnetii / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D004251:DNA Transposable Elements; D020869:Gene Expression Profiling / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057166:High-Throughput Screening Assays / Q000379:methods*; D054884:Host-Pathogen Interactions; D016254:Mutagenesis, Insertional / Q000379:methods; D010641:Phenotype; D059467:Transcriptome
chemical_list: D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins