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转录组分析表明,多药外排基因上调可保护铜绿假单胞菌免受五氯苯酚胁迫。

Transcriptome analysis reveals that multidrug efflux genes are upregulated to protect Pseudomonas aeruginosa from pentachlorophenol stress.

作者信息

Muller Jocelyn Fraga, Stevens Ann M, Craig Johanna, Love Nancy G

机构信息

Department of Civil and Environmental Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.

出版信息

Appl Environ Microbiol. 2007 Jul;73(14):4550-8. doi: 10.1128/AEM.00169-07. Epub 2007 May 25.

Abstract

Through chemical contamination of natural environments, microbial communities are exposed to many different types of chemical stressors; however, research on whole-genome responses to this contaminant stress is limited. This study examined the transcriptome response of a common soil bacterium, Pseudomonas aeruginosa, to the common environmental contaminant pentachlorophenol (PCP). Cells were grown in chemostats at a low growth rate to obtain substrate-limited, steady-state, balanced-growth conditions. The PCP stress was administered as a continuous increase in concentration, and samples taken over time were examined for physiological function changes with whole-cell acetate uptake rates (WAURs) and cell viability and for gene expression changes by Affymetrix GeneChip technology and real-time reverse transcriptase PCR. Cell viability, measured by heterotrophic plate counts, showed a moderately steady decrease after exposure to the stressor, but WAURs did not change in response to PCP. In contrast to the physiological data, the microarray data showed significant changes in the expression of several genes. In particular, genes coding for multidrug efflux pumps, including MexAB-OprM, were strongly upregulated. The upregulation of these efflux pumps protected the cells from the potentially toxic effects of PCP, allowing the physiological whole-cell function to remain constant.

摘要

通过自然环境的化学污染,微生物群落会接触到许多不同类型的化学应激源;然而,关于全基因组对这种污染物应激反应的研究却很有限。本研究检测了一种常见土壤细菌铜绿假单胞菌对常见环境污染物五氯苯酚(PCP)的转录组反应。细胞在恒化器中以低生长速率培养,以获得底物受限、稳态、平衡生长的条件。PCP应激通过浓度的持续增加来施加,对随时间采集的样本进行检测,以观察全细胞乙酸摄取率(WAURs)和细胞活力的生理功能变化,以及通过Affymetrix基因芯片技术和实时逆转录PCR检测基因表达变化。通过异养平板计数法测定的细胞活力在暴露于应激源后呈适度稳定下降,但WAURs对PCP没有反应。与生理数据相反,微阵列数据显示几个基因的表达有显著变化。特别是,编码多药外排泵(包括MexAB-OprM)的基因被强烈上调。这些外排泵的上调保护细胞免受PCP潜在的毒性作用,使生理全细胞功能保持恒定。

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