{{'Search' | translate}}
 

Incubation shaker Unitron

Company: Infors
Catalog#: Plus AJ252
Bio-protocol()
Company-protocol()
Other protocol()

Measurement of Energy-dependent Rhodamine 6G Efflux in Yeast Species
Author:
Date:
2017-08-05
[Abstract]  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. [摘要]  罗丹明6G是一种高荧光染料,常用于测定酵母膜外排泵的运输活性。 ATP结合盒转运蛋白Klr5p赋予乳酸克鲁维酵酵菌中几种不相关药物的抗性。 Pdr5p还以能量依赖的方式从完整的酵母细胞中挤出罗丹明6G(R6G)。 在2-脱氧-D-葡萄糖(糖酵解抑制剂)和R6G(线粒体ATP酶抑制剂)存在下酵母细胞的孵育导致细胞内ATP池的显着消耗(Kolaczkowski等人,1996)。 从完整的酵母细胞中可以直接测量R6G的荧光的直接测量PdF5p介导的来自完整酵母细胞的R6G的挤出。
【背景】多药物外排泵广泛分布,可在所有生物种类中找到。它们代表抗菌素耐药性的重要机制。量化外排泵活性的能力对于理解其对生理过程的贡献和评估潜在治疗药物(例如,外排抑制剂)的有效性(Blair和Piddock,2016)是必要的。外排活动测量的方法主要依赖于两种不同的机制。一些方法直接测量底物流出,即,多少基质被泵出,其他方法测量细胞内的底物分子积累,其水平然后用于间接推断流出。然而,后者由于可改变膜渗透性而改变了染料流入速率(Blair和Piddock,2016)的敏感性较差。 R6G在生长中的积累。白念珠菌细胞与ABC转运蛋白念珠菌药物抗性1(CDR1 ...

Comments