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

Date

2010-11

Authors

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

Journal Title

Journal ISSN

Volume Title

Publisher

Future Science Group

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.

Description

Keywords

Citation

Karna, S. L. R., Zogaj, X., Barker, J. R., Seshu, J., Dove, S. L., & Klose, K. E. (2010). A bacterial two-hybrid system that utilizes Gateway cloning for rapid screening of protein-protein interactions. BioTechniques, 49(5), 831-833. doi:10.2144/000113539

Department

Molecular Microbiology and Immunology
Neuroscience, Developmental and Regenerative Biology