title: Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus.
abstract: One important feature of the major opportunistic human pathogen Staphylococcus aureus is its extraordinary ability to rapidly acquire resistance to antibiotics. Genomic studies reveal that S. aureus carries many virulence and resistance genes located in mobile genetic elements, suggesting that horizontal gene transfer (HGT) plays a critical role in S. aureus evolution. However, a full and detailed description of the methodology used to study HGT in S. aureus is still lacking, especially regarding natural transformation, which has been recently reported in this bacterium. This work describes three protocols that are useful for the in vitro investigation of HGT in S. aureus: conjugation, phage transduction, and natural transformation. To this aim, the cfr gene (chloramphenicol/florfenicol resistance), which confers the Phenicols, Lincosamides, Oxazolidinones, Pleuromutilins, and Streptogramin A (PhLOPSA)-resistance phenotype, was used. Understanding the mechanisms through which S. aureus transfers genetic materials to other strains is essential to comprehending the rapid acquisition of resistance and helps to clarify the modes of dissemination reported in surveillance programs or to further predict the spreading mode in the future.
authors.affiliation: Division of Biomedical Science, Faculty of Medicine, University of Tsukuba; Department of Basic Biomedical Science, Universidad Europea de Madrid; fabio.cafini@universidadeuropea.es.
authors.forename: Fabio
authors.initials: F
authors.lastname: Cafini
authors.affiliation: Human Biology Program, School of Integrative and Global Majors, University of Tsukuba.
authors.forename: Nguyen
authors.initials: N
authors.lastname: Thi Le Thuy
authors.affiliation: Laboratory of Nosocomial Infections, Department of Bacteriology, Centro Nacional de MicrobiologÍa, Instituto de Salud Carlos III.
authors.forename: Federico
authors.initials: F
authors.lastname: Román
authors.affiliation: Division of Microbiology, Department of Medicine, School of Medicine, Universidad Complutense.
authors.forename: José
authors.initials: J
authors.lastname: Prieto
authors.affiliation: Biology of Gram-Positive Pathogens, Department of Microbiology, Institut Pasteur, Paris, France; ERL3526, CNRS, Paris, France.
authors.forename: Sarah
authors.initials: S
authors.lastname: Dubrac
authors.affiliation: Biology of Gram-Positive Pathogens, Department of Microbiology, Institut Pasteur, Paris, France; ERL3526, CNRS, Paris, France.
authors.forename: Tarek
authors.initials: T
authors.lastname: Msadek
authors.affiliation: Division of Biomedical Science, Faculty of Medicine, University of Tsukuba.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Morikawa
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics*; D003227:Conjugation, Genetic / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000187:drug effects / Q000235:genetics*; D022761:Gene Transfer, Horizontal; D005821:Genetic Techniques; D006801:Humans; D013204:Staphylococcus Phages / Q000235:genetics; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics*; D013839:Thiamphenicol / Q000031:analogs & derivatives / Q000494:pharmacology; D014161:Transduction, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; C539943:CFR protein, Staphylococcus aureus; C035534:florfenicol; D013839:Thiamphenicol