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[在有和没有gyrA突变的鼠伤寒沙门氏菌分离株中,与AcrAB/TolC系统和膜通透性相关的基因表达特征]

[Characterization of gene expression associated with both the AcrAB/TolC system and the membrane permeability, in Salmonella spp isolates with and without gyrA mutation].

作者信息

Galiana Antonio, Sánchez-Guillén Laura, Rodríguez Juan Carlos, Cremades Rosa, Santibañez Migue, Ferrari Rafaela, Ruiz-García Montserrat, López Pilar, Royo Gloria

机构信息

Juan Carlos Rodríguez, Hospital General Universitario de Alicante, Sección de Microbiología, C/ Pintor Baeza 10 - 03010 Alicante, Spain.

出版信息

Rev Esp Quimioter. 2014 Dec;27(4):239-43.

Abstract

Introduction. The marA, soxS, ramA, acrB and ompF genes have been studied in order to characterize mechanisms of AcrAB-TolC active efflux pumps and membrane permeability alterations that reduce fluoroquinolones susceptibility in Salmonella spp. Methods. Mutations in marA, soxS, ramA, acrB and ompF genes were detected, as well as their expression levels in presence and absence of ciprofloxacin, calculating the level of change between them by qPCR. Data were analysed by using SPSS 19.0. Results. No mutations in these genes were found, but both AcrAB-TolC regulatory genes and structural acrB gene expression were affected by ciprofloxacin in both mutant strains and wild type bacterial strains (WT). The activation of the marA gene in presence of drug was higher in WT strains (level of change 0.823) than in mutants strains (level of change 0.158; p=0.049). In gyrA mutants, a reduction in ompF gene expression in presence of ciprofloxacin was found (level of change -0.949 p=0.017). Conclusion. The reduction of fluoroquinolones susceptibility in Salmonella spp is a complex process, in which several different bacterial mechanisms are involved. This study has found a high difference in the degree of participation among studied mechanisms, between bacterial strains with and without gyrA mutation. Whereas WT strains activated efflux pumps especially through marA gene, mutants supressed ompF gene expression related to porins.

摘要

引言。对marA、soxS、ramA、acrB和ompF基因进行了研究,以阐明AcrAB - TolC活性外排泵的机制以及导致沙门氏菌对氟喹诺酮类药物敏感性降低的膜通透性改变。方法。检测marA、soxS、ramA、acrB和ompF基因的突变情况,以及在有和没有环丙沙星存在时它们的表达水平,通过qPCR计算它们之间的变化水平。使用SPSS 19.0分析数据。结果。在这些基因中未发现突变,但环丙沙星对突变菌株和野生型细菌菌株(WT)中的AcrAB - TolC调节基因和结构acrB基因的表达均有影响。野生型菌株在药物存在时marA基因的激活程度(变化水平0.823)高于突变菌株(变化水平0.158;p = 0.049)。在gyrA突变体中,发现环丙沙星存在时ompF基因表达降低(变化水平 - 0.949,p = 0.017)。结论。沙门氏菌对氟喹诺酮类药物敏感性的降低是一个复杂的过程,涉及多种不同的细菌机制。本研究发现,在有和没有gyrA突变的细菌菌株之间,所研究机制的参与程度存在很大差异。野生型菌株尤其通过marA基因激活外排泵,而突变体则抑制与孔蛋白相关的ompF基因表达。

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