A bacterial two-hybrid system that utilizes Gateway cloning for rapid screening of protein-protein interactions

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...

Ausführliche Beschreibung

Bibliographische Detailangaben
Veröffentlicht in:	BioTechniques. - 1991. - 49(2010), 5 vom: 15. Nov., Seite 831-3
1. Verfasser:	Karna, S L Rajasekhar (VerfasserIn)
Weitere Verfasser:	Zogaj, Xhavit, Barker, Jeffrey R, Seshu, Janakiram, Dove, Simon L, Klose, Karl E
Format:	Online-Aufsatz
Sprache:	English
Veröffentlicht:	2010
Zugriff auf das übergeordnete Werk:	BioTechniques
Schlagworte:	Journal Article Research Support, N.I.H., Extramural Bacterial Proteins Early Growth Response Protein 1 Egr1 protein, mouse Recombinant Fusion Proteins DNA-Directed RNA Polymerases EC 2.7.7.6

Beschreibung
Zusammenfassung:	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
Beschreibung:	Date Completed 24.02.2011 Date Revised 24.11.2010 published: Print Citation Status MEDLINE
ISSN:	1940-9818
DOI:	10.2144/000113539