title: A Method for Characterizing the Type III Secretion System's Contribution to Pathogenesis: Homologous Recombination to Generate Yersinia pestis Type III Secretion System Mutants.
abstract: The type III (T3S) secretion system of many gram-negative bacteria is a surface-exposed protein secretion apparatus used to directly inject bacterial effector molecules into eukaryotic cells. These effector molecules contribute to bacterial pathogenesis in many ways, and have been shown to be crucial for infectivity. Here, we describe a protocol for using homologous recombination to generate T3S system mutants to assess the role of different T3S system proteins in bacterial pathogenesis.
authors.affiliation: Department of Microbiology, University of Chicago, Chicago, IL, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Osei-Owusu
authors.affiliation: Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, Grand Forks, ND, USA. matthew.nilles@med.und.edu.
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Nilles
authors.affiliation: Department of Biomedical Sciences, University of North Dakota, 1301 North Columbia Road, Stop 9037, Grand Forks, ND, 58203, USA.
authors.forename: David S
authors.initials: DS
authors.lastname: Bradley
authors.affiliation: Department of Biomedical Sciences, University of North Dakota, 1301 North Columbia Road, Stop 9037, Grand Forks, ND, 58203, USA.
authors.forename: Travis D
authors.initials: TD
authors.lastname: Alvine
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006367:HeLa Cells; D059765:Homologous Recombination; D006801:Humans; D051379:Mice; D009154:Mutation; D010930:Plague / Q000382:microbiology; D021381:Protein Transport; D000069296:Type III Secretion Systems / Q000235:genetics* / Q000378:metabolism*; D015010:Yersinia pestis / Q000235:genetics* / Q000378:metabolism* / Q000472:pathogenicity
chemical_list: D001426:Bacterial Proteins; D000069296:Type III Secretion Systems
keywords: Homologous recombination; Pathogenesis; Secretion; Translocation; Type III secretion; Yersinia pestis