title: Expression and Purification of N-Terminally His-Tagged Recombinant Type III Secretion Proteins.
abstract: The ability to express and purify recombinant needle proteins from the Type III Secretion System (T3SS) of many gram-negative bacteria has allowed us to develop novel experimental approaches, both in vitro and in vivo, to identify unique roles for T3SS in bacterial pathogenesis. In addition, these purified needle proteins have shown to be promising immunotherapies acting as both protective antigens and adjuvants, presumably due to their immune activating properties. Here, we describe the expression and purification of recombinant T3SS needle proteins.
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
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.
authors.forename: Danielle L Jessen
authors.initials: DL
authors.lastname: Condry
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
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification*; D002845:Chromatography; D003001:Cloning, Molecular; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins; D014169:Transformation, Bacterial; D000069296:Type III Secretion Systems / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D000069296:Type III Secretion Systems
keywords: Blunt-end PCR cloning; His-tag; Protein expression; Protein purification; pET200 vector