Novel mobilizable prokaryotic two-hybrid system vectors for high-throughput protein interaction mapping in Escherichia coli by bacterial conjugation

Clarke, Paul, Ó Cuív, Páraic and O'Connell, Michael (2005) Novel mobilizable prokaryotic two-hybrid system vectors for high-throughput protein interaction mapping in Escherichia coli by bacterial conjugation. Nucleic Acids Research, 33 2: e18.1-e18.12. doi:10.1093/nar/gni011


Author Clarke, Paul
Ó Cuív, Páraic
O'Connell, Michael
Title Novel mobilizable prokaryotic two-hybrid system vectors for high-throughput protein interaction mapping in Escherichia coli by bacterial conjugation
Formatted title
Novel mobilizable prokaryotic two-hybrid system vectors for high-throughput protein interaction mapping in Escherichia coli by bacterial conjugation
Journal name Nucleic Acids Research   Check publisher's open access policy
ISSN 0305-1048
1362-4962
1362-4954
Publication date 2005-01-01
Sub-type Article (original research)
DOI 10.1093/nar/gni011
Open Access Status DOI
Volume 33
Issue 2
Start page e18.1
End page e18.12
Total pages 12
Place of publication Oxford, United Kingdom
Publisher Oxford University Press
Language eng
Formatted abstract
Since its initial description, the yeast two-hybrid (Y2H) system has been widely used for the detection and analysis of protein–protein interactions. Mating-based strategies have been developed permitting its application for automated proteomic interaction mapping projects using both exhaustive and high-throughput strategies. More recently, a number of prokaryotic two-hybrid (P2H) systems have been developed but, despite the many advantages such Escherichia coli-based systems have over the Y2H system, they have not yet been widely implemented for proteomic interaction mapping. This may be largely due to the fact that high-throughput strategies employing bacterial transformation are not as amenable to automation as Y2H mating-based strategies. Here, we describe the construction of novel conjugative P2H system vectors. These vectors carry a mobilization element of the IncPα group plasmid RP4 and can therefore be mobilized with high efficiency from an E.coli donor strain encoding all of the required transport functions in trans. We demonstrate how these vectors permit the exploitation of bacterial conjugation for technically simplified and automated proteomic interaction mapping strategies in E.coli, analogous to the mating-based strategies developed for the Y2H system.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: UQ Diamantina Institute Publications
 
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Created: Wed, 19 Mar 2014, 22:51:26 EST by Paraic O Cuiv on behalf of UQ Diamantina Institute