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KMID : 0545120130230030430
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Journal of Microbiology and Biotechnology
2013 Volume.23 No. 3 p.430 ~ p.435
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Gene Cloning and Characterization of MdeA, a Novel Multidrug Efflux Pump in Streptococcus mutans
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Kim Do-Kyun
Kim Kyoung-Hoon
Cho Eun-Ji
Joo Seoung-Je
Chung Jung-Min
Son Byoung-Yil
Yum Jong-Hwa
Kim Young-Man
Kwon Hyun-Ju
Kim Byung-Woo
Kim Tae-Hoon
Lee Eun-Woo
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Abstract
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Multidrug resistance, especially multidrug efflux mechanisms that extrude structurally unrelated cytotoxic compounds from the cell by multidrug transporters, is a serious problem and one of the main reasons for the failure of therapeutic treatment of infections by pathogenic microorganisms as well as of cancer cells. Streptococcus mutans is considered one of the primary causative agents of dental caries and periodontal disease, which comprise the most common oral diseases. A fragment of chromosomal DNA from S. mutans KCTC3065 was cloned using Escherichia coli KAM32 as host cells lacking major multidrug efflux pumps. Although E. coli KAM32 cells were very sensitive to many antimicrobial agents, the transformed cells harboring a recombinant plasmid became resistant to several structurally unrelated antimicrobial agents such as tetracycline, kanamycin, rhodamin 6G, ampicillin, acriflavine, ethidium bromide, and tetraphenylphosphonium chloride. This suggested that the cloned DNA fragment carries a gene encoding a multidrug efflux pump. Among 49 of the multidrug-resistant transformants, we report the functional gene cloning and characterization of the function of one multidrug efflux pump, namely MdeA from S. mutans, which was expressed in E. coli KAM32. Judging from the structural and biochemical properties, we concluded that MdeA is the first cloned and characterized multidrug efflux pump using the proton motive force as the energy for efflux drugs.
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KEYWORD
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Streptococcus mutans, multidrug resistance, drug efflux pump, gene cloning
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