title: A bacterial two-hybrid system that utilizes Gateway cloning for rapid screening of protein-protein interactions.
abstract: Comprehensive clone sets representing the entire genome now exist for a large number of organisms. The Gateway entry clone sets are a particularly useful means to study gene function, given the ease of introduction into any Gateway-suitable destination vector. We have adapted a bacterial two-hybrid system for use with Gateway entry clone sets, such that potential interactions between proteins encoded within these clone sets can be determined by new destination vectors. We show that utilizing the Gateway clone sets for Francisella tularensis and Vibrio cholerae, known interactions between F. tularensis IglA and IglB and V. cholerae VipA and VipB could be confirmed with these destination vectors. Moreover, the introduction of unique tags into each vector allowed for visualization of the expressed hybrid proteins via Western immunoblot. This Gateway-suitable bacterial two-hybrid system provides a new tool for rapid screening of protein-protein interactions.
authors.affiliation: South Texas Center for Emerging Infectious Diseases and Department of Biology, University of Texas San Antonio, San Antonio, TX 78249, USA.
authors.forename: S L Rajasekhar
authors.initials: SL
authors.lastname: Karna
authors.forename: Xhavit
authors.initials: X
authors.lastname: Zogaj
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Barker
authors.forename: Janakiram
authors.initials: J
authors.lastname: Seshu
authors.forename: Simon L
authors.initials: SL
authors.lastname: Dove
authors.forename: Karl E
authors.initials: KE
authors.lastname: Klose
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D051766:Early Growth Response Protein 1 / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005604:Francisella tularensis / Q000737:chemistry / Q000235:genetics; D005822:Genetic Vectors; D051379:Mice; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D020798:Two-Hybrid System Techniques; D014734:Vibrio cholerae / Q000737:chemistry / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D051766:Early Growth Response Protein 1; C486109:Egr1 protein, mouse; D011993:Recombinant Fusion Proteins; D012321:DNA-Directed RNA Polymerases