Functional gene cloning and characterization of MdeA, a multidrug efflux pump from Staphylococcus aureus

Youichi Yamada, Sumiko Shiota, Tohru Mizushima, Teruo Kuroda, Tomofusa Tsuchiya

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)801-804
Number of pages4
JournalBiological and Pharmaceutical Bulletin
Volume29
Issue number4
DOIs
Publication statusPublished - Apr 2006

Fingerprint

Staphylococcus aureus
Organism Cloning
DNA
Ethidium
Genes
Escherichia coli
Anti-Infective Agents
Norfloxacin
Daunorubicin
Drug Resistance
Doxorubicin
Plasmids
Growth
Pharmaceutical Preparations
HOE 33342
tetraphenylphosphonium

Keywords

  • Drug resistance
  • MdeA
  • Multidrug efflux pump
  • Staphylococcus aureus

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Functional gene cloning and characterization of MdeA, a multidrug efflux pump from Staphylococcus aureus. / Yamada, Youichi; Shiota, Sumiko; Mizushima, Tohru; Kuroda, Teruo; Tsuchiya, Tomofusa.

In: Biological and Pharmaceutical Bulletin, Vol. 29, No. 4, 04.2006, p. 801-804.

Research output: Contribution to journalArticle

Yamada, Youichi ; Shiota, Sumiko ; Mizushima, Tohru ; Kuroda, Teruo ; Tsuchiya, Tomofusa. / Functional gene cloning and characterization of MdeA, a multidrug efflux pump from Staphylococcus aureus. In: Biological and Pharmaceutical Bulletin. 2006 ; Vol. 29, No. 4. pp. 801-804.
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