Functional expression of candida albicans drug efflux pump Cdr1p in a Saccharomyces cerevisiae strain deficient in membrane transporters
Functional expression of candida albicans drug efflux pump Cdr1p in a Saccharomyces cerevisiae strain deficient in membrane transporters
复制标题
DOI：
10.1128/aac.45.12.3366-3374.2001
复制
发表时间：
2001-12-01
影响因子：
4.9
通讯作者：
Cannon, RD
中科院分区：
文献类型：
--
作者：
Nakamura, K;Niimi, M;Cannon, RD
对单个白色念珠菌质膜药物流出泵的转运功能的分析受到多种内源性转运蛋白的阻碍。我们从酿酒酵母突变体 (AD1-8u(-)) 的基因组 PDR5 位点稳定表达了白色念珠菌 Cdr1p，它是与多重耐药表型有关的主要泵，其中 ATP 结合盒 (ABC) 家族的 7 个主要转运蛋白已被删除。 Cdr1p 在酿酒酵母 PDR5 启动子的控制下并由酿酒酵母 Pdr1p 转录调节因子突变 pdr1-3 驱动，通过 mRNA 转录水平的增加、核苷三磷酸酶活性水平的增加以及质膜组分中的免疫检测来证明。酿酒酵母 AD1-8u(-) 对唑类抗真菌药物过敏（氟康唑抑制 80% 细胞的 MIC [MIC(80)s] 为 0.625 mug/ml，
Analysis of the transport functions of individual Candida albicans plasma membrane drug efflux pumps is hampered by the multitude of endogenous transporters. We have stably expressed C. albicans Cdr1p, the major pump implicated in multiple-drug-resistance phenotypes, from the genomic PDR5 locus in a Saccharomyces cerevisiae mutant (AD1-8u(-)) from which seven major transporters of the ATP-binding cassette (ABC) family have been deleted. High-level expression of Cdr1p, under the control of the S. cerevisiae PDR5 promoter and driven by S. cerevisiae Pdr1p transcriptional regulator mutation pdr1-3, was demonstrated by increased levels of mRNA transcription, increased levels of nucleoside triphosphatase activity, and immunodetection in plasma membrane fractions. S. cerevisiae AD1-8u(-) was hypersensitive to azole antifungals (the MICs at which 80% of cells were inhibited [MIC(80)s] were 0.625 mug/ml for fluconazole,