AmrB对嗜水气单胞菌氨基糖苷类抗生素耐药性的影响

doi:10.3969/j.issn.1008-0864.2013.03.17

›› 2013, Vol. 15 ›› Issue (3): 123-128.DOI: 10.3969/j.issn.1008-0864.2013.03.17

AmrB对嗜水气单胞菌氨基糖苷类抗生素耐药性的影响

张丹凤1,陈国平2,张国广1,黄家福1,华秀婷1,陈阳1

(1.漳州师范学院生物科学与技术系, 福建漳州 363000|2.漳州市农业局种植业管理站, 福建漳州 363000)

收稿日期:2013-02-28 修回日期:2013-03-28 出版日期:2013-06-15 发布日期:2013-06-15
作者简介:张丹凤,副教授,博士,主要从事微生物学研究。E-mail:zh_danfeng@163.com
基金资助:
福建省自然科学基金项目(2010J05088);漳州市自然科学基金项目(zz2012J05)资助。

The Effect of AmrB on Aeromonas hydrophila Aminoglycosides Resistance

ZHANG Dan-feng1, CHEN Guo-ping2, ZHANG Guo-guang1, HUANG Jia-fu1, HUA Xiu-ting1, CHEN Yang1

(1.Department of Biological Sciences and Biotechnology, Zhangzhou Normal University, Fujian Zhangzhou, 363000|2.Planting Management Station, Zhangzhou Agriculture Bureau, Fujian Zhangzhou 363000, China)

Received:2013-02-28 Revised:2013-03-28 Online:2013-06-15 Published:2013-06-15

摘要/Abstract

摘要：

AmrB是药物外排系统的重要成员,探讨AmrB与嗜水气单胞菌氨基糖苷类抗生素耐药性的关系,对研究嗜水气单胞菌耐药机制有重要意义。将amrB基因克隆至pET-28a载体上,双酶切和测序验证得到重组质粒amrB-pET-28a。对amrB-pET-28a/BL21进行诱导表达和镍柱纯化,制备得到效价为1∶6 400的兔抗AmrB抗血清。诱导获得嗜水气单胞菌AH1卡那霉素、链霉素和庆大霉素耐药菌株,Western blotting分析AmrB在耐药菌株中的表达量。Phoretix 1D和SPSS分析表明,AmrB的表达量在卡那霉素、链霉素和庆大霉素耐药菌株中显著地高于对照菌嗜水气单胞菌AH1,分别是对照的2.16、2.65和2.13倍,提示AmrB在嗜水气单胞菌对氨基糖苷类抗生素耐药过程中起重要作用,这将有助于了解嗜水气单胞菌耐药的分子机制。

关键词: 嗜水气单胞菌;AmrB;耐药;氨基糖苷类抗生素

Abstract:

AmrB is a crucial member of drug efflux pump. Investigating the relationship between AmrB and the aminoglycosides resistance of Aeromonas hydrophila is supposed to be important for understanding the mechanism of its drug resistance. amrB gene was cloned to pET-28a vector and confirmed by double digestion and sequencing, thus obtained amrB-pET-28a. amrB-pET-28a/BL21 was induced by IPTG to obtain AmrB protein then purified by Ni resin. New Zealand rabbit was immunized by AmrB to get AmrB serum with a titer of 1∶6 400. Kanamycin, streptomycin and gentamycin resistant strains of Aeromonas hydrophila AH1 were screened by 10 serial culture with 1/2 MIC of corresponding antibiotic. Western blotting was performed to analyze the expression of AmrB in drug resistant strains. After analysis of Western blotting by Phoretix 1D software and SPSS software, the expression of AmrB in kanamycin, streptomycin and gentamycin resistant strains was significantly 2.16,2.65 and 2.13 folds higher than control strain, respectively. AmrB plays an important role in the aminoglycosides resistance of Aeromonas hydrophila AH1, which may offer an opportunity to further explore the molecular mechanisms within.

Key words: Aeromonas hydrophila, AmrB, drug resistance, aminoglycosides

中图分类号:

Q816

张丹凤1,陈国平2,张国广1,黄家福1,华秀婷1,陈阳1 . AmrB对嗜水气单胞菌氨基糖苷类抗生素耐药性的影响[J]. , 2013, 15(3): 123-128.

ZHANG Dan-feng1, CHEN Guo-ping2, ZHANG Guo-guang1, HUANG Jia-fu1, HUA Xiu-ting1,. The Effect of AmrB on Aeromonas hydrophila Aminoglycosides Resistance[J]. , 2013, 15(3): 123-128.

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AmrB对嗜水气单胞菌氨基糖苷类抗生素耐药性的影响

The Effect of AmrB on Aeromonas hydrophila Aminoglycosides Resistance

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