Functional gene cloning and characterization of MdeA, a multidrug efflux pump from Staphylococcus aureus
Functional gene cloning and characterization of MdeA, a multidrug efflux pump from Staphylococcus aureus
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DOI：
10.1248/bpb.29.801
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发表时间：
2006-04-01
影响因子：
2
通讯作者：
Tsuchiya, T
中科院分区：
文献类型：
--
作者：
Yamada, Y;Shiota, S;Tsuchiya, T
以药物敏感性大肠杆菌KAM 32为宿主，从金黄色葡萄球菌N315的染色体DNA中克隆了赋予药物抗性的DNA片段。虽然E.大肠杆菌KAM 32细胞对多种抗菌药物敏感，携带携带DNA区域的重组质粒的转化细胞对几种结构无关的抗菌药物产生抗性，如氯化四苯基磷、Hoechst 33342和诺氟沙星。这些结果表明，克隆的DNA片段携带编码多药外排泵的基因。我们对克隆的DNA进行了部分核苷酸序列测定，发现其中含有mdeA基因。用mdeA基因转化的大肠杆菌细胞显示Hoechst 33342的外排活性。另一方面，S.用mdeA转化的金黄色葡萄球菌细胞显示对阿霉素、柔红霉素、氯化四苯基磷、Hoechst 33342、溴化乙锭和罗丹明6G的抗性升高。在转化的S.金黄色。我们发现mdeA基因在正常生长条件下在S. aureus N315。
A DNA fragment conferring drug resistance was cloned from the chromosomal DNA of Staphylococcus aureus N315 using a drug hypersensitive Escherichia coli KAM32 as the host. Although E. coli KAM32 cells were sensitive to many antimicrobial agents, transformed cells harboring a recombinant plasmid carrying the DNA region became resistant to several structurally unrelated antimicrobial agents, such as tetraphenylphosphonium chloride, Hoechst 33342 and norfloxacin. These results suggest that the cloned DNA fragment carries a gene(s) encoding a multidrug efflux pump. We partially determined the nucleotide sequence of the cloned DNA and found the mdeA gene within it. The E. coli cells transformed with the mdeA gene showed efflux activity of Hoechst 33342. On the other hand, S. aureus cells transformed with mdeA showed elevated resistance to doxorubicin, daunorubicin, tetraphenylphosphonium chloride, Hoechst 33342, ethidium bromide and rhodamine 6G. Elevated energy-dependent efflux of ethidium was observed with transformed S. aureus. We found that the mdeA gene was expressed under normal growth conditions in S. aureus N315.