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Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence
We present a fluorometric method for determining ABC transporter activity in the pathogenic fungus C. albicans during different growth phases and in response to glucose. The carbocyanine dye diS-C(3)(3) was previously used to monitor plasma membrane potentials and test the influence of surface-activ...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2015
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673308/ https://www.ncbi.nlm.nih.gov/pubmed/26696990 http://dx.doi.org/10.3389/fmicb.2015.01382 |
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author | Szczepaniak, Joanna Łukaszewicz, Marcin Krasowska, Anna |
author_facet | Szczepaniak, Joanna Łukaszewicz, Marcin Krasowska, Anna |
author_sort | Szczepaniak, Joanna |
collection | PubMed |
description | We present a fluorometric method for determining ABC transporter activity in the pathogenic fungus C. albicans during different growth phases and in response to glucose. The carbocyanine dye diS-C(3)(3) was previously used to monitor plasma membrane potentials and test the influence of surface-active compounds in membrane polarization. We used diS-C(3)(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-C(3)(3) results by standard RT-PCR and Western blotting. |
format | Online Article Text |
id | pubmed-4673308 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-46733082015-12-22 Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence Szczepaniak, Joanna Łukaszewicz, Marcin Krasowska, Anna Front Microbiol Microbiology We present a fluorometric method for determining ABC transporter activity in the pathogenic fungus C. albicans during different growth phases and in response to glucose. The carbocyanine dye diS-C(3)(3) was previously used to monitor plasma membrane potentials and test the influence of surface-active compounds in membrane polarization. We used diS-C(3)(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-C(3)(3) results by standard RT-PCR and Western blotting. Frontiers Media S.A. 2015-12-09 /pmc/articles/PMC4673308/ /pubmed/26696990 http://dx.doi.org/10.3389/fmicb.2015.01382 Text en Copyright © 2015 Szczepaniak, Łukaszewicz and Krasowska. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Szczepaniak, Joanna Łukaszewicz, Marcin Krasowska, Anna Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence |
title | Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence |
title_full | Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence |
title_fullStr | Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence |
title_full_unstemmed | Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence |
title_short | Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence |
title_sort | estimation of candida albicans abc transporter behavior in real-time via fluorescence |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673308/ https://www.ncbi.nlm.nih.gov/pubmed/26696990 http://dx.doi.org/10.3389/fmicb.2015.01382 |
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