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BEEP: An assay to detect bio-energetic and envelope permeability alterations in Pseudomonas aeruginosa.

Abstract：

:We developed an effective and rapid assay to detect both bio-energetic and envelope permeability (BEEP) alterations of Pseudomonas aeruginosa. The assay is based on quantification of extracellular ATP in bacterial cultures using luciferase as a reporter. To demonstrate the validity of our assay we conducted a biased screen of a transposon insertion library in P. aeruginosa strain PAO1 in order to expedite the isolation of mutants with defects in bioenergetic pathways. We successfully isolated insertion mutants that were reduced for extracellular ATP accumulation and identified the corresponding mutations that caused the phenotype. Most of the genes identified from this analysis were associated with energy metabolism and several appeared to be potentially novel bioenergetic targets. In addition, we show that treatment of P. aeruginosa strain PAO1 with antibiotics that disrupt the bacterial cell envelope leads to greater extracellular ATP accumulation. In summary, increases in extracellular ATP accumulation above wild type levels indicated a perturbation of membrane permeability while decreases in extracellular ATP accumulation indicated defects in bioenergetics.

journal_name

J Microbiol Methods

journal_title

Journal of microbiological methods

authors

Suh SJ,Shuman J,Carroll LP,Silo-Suh L

doi

10.1016/j.mimet.2016.04.009

subject

Has Abstract

pub_date

2016-06-01 00:00:00

pages

81-6

eissn

0167-7012

issn

1872-8359

pii

S0167-7012(16)30069-0

journal_volume

125

pub_type

杂志文章

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