Functional cloning and expression of emeA, and characterization of EmeA, a multidrug efflux pump from Enterococcus faecalis

Eun Woo Lee, Jing Chen, Nazmul Huda, Teruo Kuroda, Tohru Mizushima, Tomofusa Tsuchiya

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

A fragment of chromosomal DNA from Enterococcus faecalis ATCC 29212 was cloned using Escherichia coli KAM32 host cells lacking major multidrug efflux pumps. E. coli KAM32 cells were sensitive to many antimicrobial agents, and the transformed cells harboring a recombinant plasmid became resistant to several structurally unrelated antimicrobial agents such as tetraphenylphosphonium chloride, 4′,6-diamidino-2-phenylindole (DAPI), Hoechst 33342, acriflavine, benzalkonium chloride, norfloxacin and ethidium bromide. This suggests that the cloned DNA fragment carries a gene(s) encoding a multidrug efflux pump. Determination of the nucleotide sequence of the cloned DNA revealed a gene designated as emeA. The transformed E. coli cells showed efflux activity of several antimicrobial agents such as DAPI, Hoechst 33342 and acriflavine. Efflux of DAPI via EmeA was strongly inhibited by reserpine.

Original languageEnglish (US)
Pages (from-to)266-270
Number of pages5
JournalBiological and Pharmaceutical Bulletin
Volume26
Issue number2
DOIs
StatePublished - Feb 1 2003
Externally publishedYes

Fingerprint

Enterococcus faecalis
Organism Cloning
Anti-Infective Agents
3,6-diamino-10-methylacridinium
Escherichia coli
DNA
Acriflavine
Benzalkonium Compounds
Norfloxacin
Ethidium
Reserpine
Genes
Plasmids
2-phenylindole
HOE 33342

Keywords

  • Drug efflux
  • Drug resistance
  • Emea multidrug efflux pump
  • Enterococcus faecalis

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

Cite this

Functional cloning and expression of emeA, and characterization of EmeA, a multidrug efflux pump from Enterococcus faecalis. / Lee, Eun Woo; Chen, Jing; Huda, Nazmul; Kuroda, Teruo; Mizushima, Tohru; Tsuchiya, Tomofusa.

In: Biological and Pharmaceutical Bulletin, Vol. 26, No. 2, 01.02.2003, p. 266-270.

Research output: Contribution to journalArticle

Lee, Eun Woo ; Chen, Jing ; Huda, Nazmul ; Kuroda, Teruo ; Mizushima, Tohru ; Tsuchiya, Tomofusa. / Functional cloning and expression of emeA, and characterization of EmeA, a multidrug efflux pump from Enterococcus faecalis. In: Biological and Pharmaceutical Bulletin. 2003 ; Vol. 26, No. 2. pp. 266-270.
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