Efflux pump gene hefA of Helicobacter pylori plays an important role in multidrug resistance
Efflux pump gene hefA of Helicobacter pylori plays an important role in multidrug resistance作者机构：Department of Gastroenterology the Second Affiliated Hospital of Zhengzhou University Zhengzhou 450014 Henan Province China Department of Pathology and Molecular Medicine McMaster University Hamilton Ontario L8N 4A6 Canada
出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)
年 卷 期：2008年第14卷第33期
页 面：5217-5222页
核心收录：
学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学]
主 题：Efflux pump Helicobacter pylori Multidrug resistance Fluorescence real-time quantitative PCR Knockout mutant
摘 要：AIM: To determine whether efflux systems contribute to multidrug resistance of H pylori. METHODS: A chloramphenicol-induced multidrug resistance model of six susceptible H pylori strains (5 isolates and H pylori NCTC11637) was developed. Multidrug-resistant (MDR) strains were selected and the minimal inhibitory concentration (MIC) of eryth-romycin, metronidazole, penicillin G, tetracycline, and ciprofloxacin in multidrug resistant strains and their parent strains was determined by agar dilution tests. The level of mRNA expression of hefA was assessed by fluorescence real-time quantitative PCR. A H pylori LZ1026 knockout mutant (ΔH pylori LZ1026) for (puta-tive) efflux protein was constructed by inserting the kanamycin resistance cassette from pEGFP-N2 into hefA, and its susceptibility profiles to 10 antibiotics were evaluated. RESULTS: The MIC of six multidrug-resistant strains (including 5 clinical isolates and H pylori NCTC11637) increased signifi cantly (≥ 4-fold) compared with their parent strains. The expression level of hefA gene was significantly higher in the MDR strains than in their parent strains (P = 0.033). A H pylori LZ1026 mutant was successfully constructed and the ΔH pylori LZ1026 was more susceptible to four of the 10 antibiotics. All the 20 strains displayed transcripts for hefA that con-fi rmed the in vitro expression of these ***: The efflux pump gene hefA plays an important role in multidrug resistance of H pylori.
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