Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1

Anowara Begum, M. Mushfequr Rahman, Wakano Ogawa, Tohru Mizushima, Teruo Kuroda, Tomofusa Tsuchiya

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

There are six putative genes for multidrug and toxic compound extrusion (MATE) family multidrug efflux pumps in the chromosome of Vibrio cholerae. We have so far analyzed two MATE family pumps in V. cholerae non-O1 NCTC4716. Here we cloned four remaining genes for putative MATE family efflux pumps by the PCR method from this microorganism and designated them as vcmB, vcmD, vcmH and vcmN. Each one of the four genes was introduced and expressed in the drug hypersusceptible host Escherichia coli KAM32 cells. We observed elevated MICs of multiple antimicrobial agents, such as fluoroquinolones, aminoglycosides, ethidium bromide and Hoechst 33342 in the transformants. Energy-dependent efflux of substrate was observed with the transformed cells. We found that efflux activities of VcmB, VcmD and VcmH were Na+-dependent, but that of VcmN was Na+-independent. Thus, all six of the MATE family multidrug efflux pumps of V. cholerae non-O1 have been characterized. We also found that all six genes were expressed in cells of V. cholerae non-O1.

Original languageEnglish
Pages (from-to)949-957
Number of pages9
JournalMicrobiology and Immunology
Volume49
Issue number11
Publication statusPublished - 2005
Externally publishedYes

Fingerprint

Vibrio cholerae non-O1
Poisons
Organism Cloning
Genes
Vibrio cholerae
Ethidium
Fluoroquinolones
Aminoglycosides
Anti-Infective Agents
Chromosomes
Escherichia coli
Polymerase Chain Reaction
Pharmaceutical Preparations

Keywords

  • MATE family
  • Multidrug efflux pump
  • V. cholerae non-O1

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Microbiology (medical)
  • Microbiology

Cite this

Begum, A., Rahman, M. M., Ogawa, W., Mizushima, T., Kuroda, T., & Tsuchiya, T. (2005). Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1. Microbiology and Immunology, 49(11), 949-957.

Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1. / Begum, Anowara; Rahman, M. Mushfequr; Ogawa, Wakano; Mizushima, Tohru; Kuroda, Teruo; Tsuchiya, Tomofusa.

In: Microbiology and Immunology, Vol. 49, No. 11, 2005, p. 949-957.

Research output: Contribution to journalArticle

Begum, A, Rahman, MM, Ogawa, W, Mizushima, T, Kuroda, T & Tsuchiya, T 2005, 'Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1', Microbiology and Immunology, vol. 49, no. 11, pp. 949-957.
Begum A, Rahman MM, Ogawa W, Mizushima T, Kuroda T, Tsuchiya T. Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1. Microbiology and Immunology. 2005;49(11):949-957.
Begum, Anowara ; Rahman, M. Mushfequr ; Ogawa, Wakano ; Mizushima, Tohru ; Kuroda, Teruo ; Tsuchiya, Tomofusa. / Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1. In: Microbiology and Immunology. 2005 ; Vol. 49, No. 11. pp. 949-957.
@article{68812bb79354442a90923a40bb3cde8e,
title = "Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1",
abstract = "There are six putative genes for multidrug and toxic compound extrusion (MATE) family multidrug efflux pumps in the chromosome of Vibrio cholerae. We have so far analyzed two MATE family pumps in V. cholerae non-O1 NCTC4716. Here we cloned four remaining genes for putative MATE family efflux pumps by the PCR method from this microorganism and designated them as vcmB, vcmD, vcmH and vcmN. Each one of the four genes was introduced and expressed in the drug hypersusceptible host Escherichia coli KAM32 cells. We observed elevated MICs of multiple antimicrobial agents, such as fluoroquinolones, aminoglycosides, ethidium bromide and Hoechst 33342 in the transformants. Energy-dependent efflux of substrate was observed with the transformed cells. We found that efflux activities of VcmB, VcmD and VcmH were Na+-dependent, but that of VcmN was Na+-independent. Thus, all six of the MATE family multidrug efflux pumps of V. cholerae non-O1 have been characterized. We also found that all six genes were expressed in cells of V. cholerae non-O1.",
keywords = "MATE family, Multidrug efflux pump, V. cholerae non-O1",
author = "Anowara Begum and Rahman, {M. Mushfequr} and Wakano Ogawa and Tohru Mizushima and Teruo Kuroda and Tomofusa Tsuchiya",
year = "2005",
language = "English",
volume = "49",
pages = "949--957",
journal = "Microbiology and Immunology",
issn = "0385-5600",
publisher = "Center for Academic Publications Japan",
number = "11",

}

TY - JOUR

T1 - Gene cloning and characterization of four MATE family multidrug efflux pumps from Vibrio cholerae non-O1

AU - Begum, Anowara

AU - Rahman, M. Mushfequr

AU - Ogawa, Wakano

AU - Mizushima, Tohru

AU - Kuroda, Teruo

AU - Tsuchiya, Tomofusa

PY - 2005

Y1 - 2005

N2 - There are six putative genes for multidrug and toxic compound extrusion (MATE) family multidrug efflux pumps in the chromosome of Vibrio cholerae. We have so far analyzed two MATE family pumps in V. cholerae non-O1 NCTC4716. Here we cloned four remaining genes for putative MATE family efflux pumps by the PCR method from this microorganism and designated them as vcmB, vcmD, vcmH and vcmN. Each one of the four genes was introduced and expressed in the drug hypersusceptible host Escherichia coli KAM32 cells. We observed elevated MICs of multiple antimicrobial agents, such as fluoroquinolones, aminoglycosides, ethidium bromide and Hoechst 33342 in the transformants. Energy-dependent efflux of substrate was observed with the transformed cells. We found that efflux activities of VcmB, VcmD and VcmH were Na+-dependent, but that of VcmN was Na+-independent. Thus, all six of the MATE family multidrug efflux pumps of V. cholerae non-O1 have been characterized. We also found that all six genes were expressed in cells of V. cholerae non-O1.

AB - There are six putative genes for multidrug and toxic compound extrusion (MATE) family multidrug efflux pumps in the chromosome of Vibrio cholerae. We have so far analyzed two MATE family pumps in V. cholerae non-O1 NCTC4716. Here we cloned four remaining genes for putative MATE family efflux pumps by the PCR method from this microorganism and designated them as vcmB, vcmD, vcmH and vcmN. Each one of the four genes was introduced and expressed in the drug hypersusceptible host Escherichia coli KAM32 cells. We observed elevated MICs of multiple antimicrobial agents, such as fluoroquinolones, aminoglycosides, ethidium bromide and Hoechst 33342 in the transformants. Energy-dependent efflux of substrate was observed with the transformed cells. We found that efflux activities of VcmB, VcmD and VcmH were Na+-dependent, but that of VcmN was Na+-independent. Thus, all six of the MATE family multidrug efflux pumps of V. cholerae non-O1 have been characterized. We also found that all six genes were expressed in cells of V. cholerae non-O1.

KW - MATE family

KW - Multidrug efflux pump

KW - V. cholerae non-O1

UR - http://www.scopus.com/inward/record.url?scp=28044441589&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=28044441589&partnerID=8YFLogxK

M3 - Article

C2 - 16301805

AN - SCOPUS:28044441589

VL - 49

SP - 949

EP - 957

JO - Microbiology and Immunology

JF - Microbiology and Immunology

SN - 0385-5600

IS - 11

ER -