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Estimation of Candida albicans ABC transporter behaviour in real-time via fluorescence

By Joanna eSzczepaniak, Marcin eŁukaszewicz and Anna eKrasowska

Abstract

We present a fluorometric method for determining ABC transporter activity in the pathogenic fungus Candida albicans during different growth phases and in response to glucose. The carbocyanine dye diS-C3(3) was previously used to monitor plasma membrane potentials and test the influence of surface-active compounds in membrane polarization. We used diS-C3(3) to show changes in fluorescence kinetics that reflect changes in the activity of ABC transporters in C. albicans growth. Cdr1-GFP fluorescence, revealed that Cdr1p relocates to the inside of the cell after the early-log growth phase. Addition of glucose to the cell suspension resulted in Cdr1p transporter expression in the CDR2-knockout strain. We confirmed the diS-C3(3) results by standard RT-PCR and Western blotting

Topics: Candida albicans, CDR, diS-C3(3) fluorescence, protein localization imaging, kinetics of efflux pump activity, Microbiology, QR1-502
Publisher: Frontiers Media S.A.
Year: 2015
DOI identifier: 10.3389/fmicb.2015.01382
OAI identifier: oai:doaj.org/article:20a9955431ef4219a28016a5e5b5c4d2
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