Gene cloning and characterization of VcrM, a Na+-coupled multidrug efflux pump, from Vibrio cholerae non-O1
Gene cloning and characterization of VcrM, a Na+-coupled multidrug efflux pump, from Vibrio cholerae non-O1
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DOI：
10.1111/j.1348-0421.2003.tb03379.x
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发表时间：
2003-01-01
影响因子：
2.6
通讯作者：
Tsuchiya, T
中科院分区：
文献类型：
--
作者：
Huda, MN;Chen, J;Tsuchiya, T
我们从非O 1型霍乱弧菌染色体上克隆了一个耐药基因片段。该片段的核苷酸序列分析揭示了编码由445个氨基酸残基组成的蛋白质的单个开放阅读框的存在。我们将该基因命名为vcrM。推导的氨基酸序列的VcrM的亲水性分析表明存在12个跨膜段。树状图显示VcrM是多药外排泵MATE家族内DinF亚家族的成员。克隆的vcrM基因在药物敏感的大肠杆菌KAM 32细胞中的表达使其对吖啶黄、4 '，6-二脒基-2-苯基吲哚、Hoechst 33342、罗丹明6 G、四苯基氯化磷（TPPCI）和溴化乙锭产生抗性。由于VcrM的外排吖啶黄依赖于Na+或Li+。此外，当TPPCI加入Na+负载的细胞时，用VcrM观察到Na+流出。因此，我们得出结论，VcrM是一个Na+/药物antiporter型多药外排泵。
We cloned a DNA fragment responsible for drug resistance from chromosome of Vibrio cholerae non-O1. Nucleotide sequence analysis of this fragment revealed the presence of a single open reading frame encoding a protein consisting of 445 amino acid residues. We designated the gene as vcrM. Hydropathy analysis of the deduced amino acid sequence of VcrM suggests the presence of 12 trans-membrane segments. A dendrogram showed that VcrM is a member of the DinF-subfamily within the MATE family of multidrug efflux pumps. Expression of the cloned vcrM gene in drug-hypersensitive Escherichia coli KAM32 cells made them resistant to acriflavine, 4', 6-diamidino-2-phenylindole, Hoechst 33342, rhodamine 6G, tetraphenylphosphonium chloride (TPPCI) and ethidium bromide. Efflux of acriflavine due to VcrM was dependent on Na+ or Li+. Moreover, Na+ efflux was observed with VcrM when TPPCI was added to Na+-loaded cells. Therefore, we conclude that VcrM is a Na+/drug antiporter-type multidrug efflux pump.