[一种rnd型多药外排泵在华奎中根菌7653R中的表型表征及表达调控]。

微生物学报 Pub Date : 2016-12-04

Ziling Liu, Jieli Peng, Youguo Li

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引用次数: 0

摘要

目的:研究华葵中根菌7653R中MCHK_0866和MCHK_0867编码的RND家族外排泵的功能。方法:分析基因组中靶基因的遗传组织。通过测定OD600来观察生长变化。用最小抑菌浓度检测药物敏感性;RT-PCR检测靶基因的相对转录水平。通过细菌单杂交系统验证了外排泵的转录调控作用。结果:MCHK_0866和MCHK_0867编码的蛋白形成了RND家族外排泵。破坏外排泵后，生长曲线OD600降低，对萘啶酸、四环素和SDS的敏感性增强。钠地酸处理后基因相对转录水平升高。同时下游基因MCHK_0869属于TetR家族转录因子，其表达产物可与MCHK_0867的启动子区相互作用。结论:外排泵可能与钠酸的转运有关，影响根瘤菌的自由生长。据推测，这种泵是由下游的局部转录因子调节的。

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[Characterization of phenotype and expression regulation of an RND-type multidrug efflux pump in Mesorhizobium huakuii 7653R].

Objective: To study the function of an RND family efflux pump encoded by MCHK_0866 and MCHK_0867 in Mesorhizobium huakuii 7653R.

Methods: Genetic organization of target genes was analyzed in genome. The change of growth was observed by measuring OD600. Drug sensitivity was detected by minimal inhibitory concentrations; relative transcription level of target genes was measured by RT-PCR. Transcript regulation of the efflux pump was validated by bacterial one-hybrid system.

Results: Proteins encoded by MCHK_0866 and MCHK_0867 formed an RND family efflux pump. The OD600 of growth curve reduced and it showed more sensitivity to nalidixic acid, tetracycline and SDS after disrupting the efflux pump. Genes relative transcription level increased in response to nalidixic acid treatment. Meanwhile, the downstream gene MCHK_0869 belongs to TetR family transcription factor and its expression product can interact with the promoter region of MCHK_0867.

Conclusion: The efflux pump is possibly associated with the transportation of nalidixic acid and affects rhizobial free-living growth. The pump is putatively regulated by a downstream local transcription factor.

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来源期刊

微生物学报

自引率

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发文量

7960

期刊介绍： Acta Microbiologica Sinica(AMS) is a peer-reviewed monthly (one volume per year)international journal,founded in 1953.It covers a wide range of topics in the areas of general and applied microbiology.The journal publishes original papers,reviews in microbiological science,and short communications describing unusual observations. Acta Microbiologica Sinica has been indexed in Index Copernicus (IC),Chemical Abstract (CA),Excerpt Medica Database (EMBASE),AJ of Viniti (Russia),Biological Abstracts (BA),Chinese Science Citation Database (CSCD),China National Knowledge Infrastructure(CNKI),Institute of Scientific and Technical Information of China(ISTIC),Chinese Journal Citation Report(CJCR),Chinese Biological Abstracts,Chinese Pharmaceutical Abstracts,Chinese Medical Abstracts and Chinese Science Abstracts.