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Construction of a bacterial autoinducer detection system in mammalian cells

External protocol Created on 09 Apr 2014

Authors

Erin K. Shiner, Sheila Reddy, Cody Timmons, Guigen Li, Simon C. Williams, Kendra P. Rumbaugh

Summary

Quorum sensing (QS) is a cell density-dependent signaling system used by bacteria to coordinate gene expression within a population. QS systems in Gram negative bacteria consist of transcription factors of the LuxR family and their acyl homoserine lactone (AHL) ligands. We describe here a method for examining QS signaling systems in mammalian cells that uses engineered LuxR-type proteins from the opportunistic pathogen, Pseudomonas aeruginosa, which can function as AHL-dependent transcription factors. The engineered proteins respond to their cognate ligands and display sequence specific DNA binding properties. This system has several potential biotechnological and biological applications. It may be used to characterize any LuxR-type protein, screen animal and plant cell extracts or exudates for compounds that mimic or interfere with AHL signaling or to screen different cell types for AHL inactivating activities.

Further details

The protocol was imported from Springer Protocols. To see the entire protocol, click on the source link, or download the pdf here.

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