Jun 12, 2018

Biolog phenotype microarray: a tool for the identification of multidrug resistance efflux pumps inducers

BioRxiv : the Preprint Server for Biology
Paula BlancoJose L Martinez

Abstract

Overexpression of multidrug resistance efflux pumps is a relevant mechanism of antibiotic resistance for bacterial pathogens. These systems use to present low levels of basal expression. However, they can be induced by environmental signals or stresses which can lead to situations of phenotypic induced resistance. In contrast to efflux pumps substrates, inducers of these systems have not been thoroughly studied. In this work, we have applied a novel high-throughput methodology in order to identify inducer molecules of the Stenotrophomonas maltophilia SmeVWX and SmeYZ efflux pumps. To that goal, bioreporters in which the expression of the yellow fluorescent protein is linked to the activity of either the smeVWX or the smeYZ promoters were developed and used for the screening of potential inducers of the expression of these efflux pumps using Biolog phenotype microarrays. Confirmation of induction was carried out measuring YFP production along the bacterial growth and by flow cytometry; mRNA levels of smeV and smeY were also determined by real-time RT-PCR after exposure to the selected compounds. Among the 144 tested compounds, iodoacetate, clioquinol (5-chloro-7-iodo-8-hydroxyquinoline) and sodium selenite were found to be smeVW...Continue Reading

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Mentioned in this Paper

Real-Time Polymerase Chain Reaction
Flow Cytometry
Erythromycin
Boric acid
Clioquinol
Antibiotic throat preparations
Antifungal Antibiotics, Topical
Environment
Iodoacetates
Promoter

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