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

S. L.Rajasekhar Karna, Xhavit Zogaj, Jeffrey R. Barker, Janakiram Seshu, Simon L. Dove, Karl E. Klose

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)831-833
Number of pages3
JournalBioTechniques
Volume49
Issue number5
DOIs
StatePublished - Nov 2010

Keywords

  • Bacterial two-hybrid
  • Francisella
  • Gateway cloning
  • Protein-protein interaction
  • Vibrio

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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