TY  - JOUR
TI  - Measurement of Energy-dependent Rhodamine 6G Efflux in Yeast Species
AU  - Gbelska, Yvetta  
AU  - Toth Hervay, Nora 
AU  - Dzugasova, Vladimira 
AU  - Konecna, Alexandra 
VL  - 7
IS  - 15
PY  - 2017
DA  - 2017/08/05
SP  - e2428
C1  - Bio-protocol 2017;7:e2428
DO  - 10.21769/BioProtoc.2428
UR  - https://doi.org/10.21769/BioProtoc.2428
AB  - Rhodamine 6G is a highly fluorescent dye often used to determine the transport activity of yeast membrane efflux pumps. The ATP-binding cassette transporter KlPdr5p confers resistance to several unrelated drugs in Kluyveromyces lactis. KlPdr5p also extrudes rhodamine 6G (R6G) from intact yeast cells in an energy-dependent manner. Incubation of yeast cells in the presence of 2-deoxy-D-glucose (inhibitor of glycolysis) and R6G (mitochondrial ATPase inhibitor) leads to marked depletion of intracellular ATP pool (Kolaczkowski et al., 1996). An active KlPdr5p mediated extrusion of R6G from intact yeast cells can be followed by direct measurement of the fluorescence of extruded R6G in the assay buffer.
KW  - Rhodamine 6G
KW  - Fluorescence
KW  - Kluyveromyces lactis
KW  - ABC transporter
KW  - Transport activity assay
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False