Abstract
In Pseudomonas aeruginosa, identification of new partners of a protein of interest could give precious clues to decipher a biological process in which this protein is involved. However, genes encoding for partners of a protein of interest are unknown and frequently scattered throughout the genome. We describe herein the construction and the use of pan-genomic bacterial two-hybrid libraries to identify new partners of a protein of interest encoded by P. aeruginosa.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Karimova G, Pidoux J, Ullmann A et al (1998) A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci U S A 95:5752–5756
Ladant D, Ullmann A (1999) Bordatella pertussis adenylate cyclase: a toxin with multiple talents. Trends Microbiol 7:172–176
Battesti A, Bouveret E (2012) The bacterial two-hybrid system based on adenylate cyclase reconstitution in Escherichia coli. Methods 58(4):325–334
Dautin N, Karimova G, Ladant D (2002) Bordetella pertussis adenylate cyclase toxin: a versatile screening tool. Toxicon 40:1383–1387
Houot L, Fanni A, de Bentzmann S et al (2012) A bacterial two-hybrid genome fragment library for deciphering regulatory networks of the opportunistic pathogen Pseudomonas aeruginosa. Microbiology 158:1964–1971
Clarke L, Carbon J (1976) A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell 9:91–99
Karimova G, Ullmann A, Ladant D (2001) Protein-protein interaction between Bacillus stearothermophilus tyrosyl-tRNA synthetase subdomains revealed by a bacterial two-hybrid system. J Mol Microbiol Biotechnol 3:73–82
Handford J, Ize B, Buchanan G et al (2009) Conserved network of proteins essential for bacterial viability. J Bacteriol 191:4732–4749
Acknowledgements
The work of C. B. and S. de B. is supported by the French Cystic Fibrosis Foundation (VLM), and CNRS institutional and ANR grants: ERA-NET ADHRES 27481, PCV-ANR 27628, ANR Jeunes Chercheurs JC09-473544 and GDR3171, as well as Europathogenomics 2005–2010 REX LSHB-CT-2005512061-EPG.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
de Bentzmann, S., Bordi, C. (2014). Construction of Pseudomonas aeruginosa Two-Hybrid Libraries for High-Throughput Assays. In: Filloux, A., Ramos, JL. (eds) Pseudomonas Methods and Protocols. Methods in Molecular Biology, vol 1149. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-0473-0_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0473-0_19
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-0472-3
Online ISBN: 978-1-4939-0473-0
eBook Packages: Springer Protocols