Israel Institute for Biological Research, Ness Ziona, Israel.
BMC Genom Data. 2023 Apr 19;24(1):23. doi: 10.1186/s12863-023-01125-6.
As part of a research aiming at presenting an alternative approach for rapid determination of antimicrobial susceptibility by quantification of changes in expression levels of specific marker genes and gene sets, cultures of the virulent bacterial strain Francisella tularensis SchuS4 were grown in the presence of inhibitory/sub-inhibitory concentrations of either ciprofloxacin or doxycycline and their transcriptomic profiles were elucidated using differential expression analysis followed by functional annotation.
RNA sequencing was performed to identify differentially expressed genes (DEGs) in response to exposure of F. tularensis SchuS4 to either ciprofloxacin or doxycycline, the antibiotics of choice for Tularemia therapy. Accordingly, RNA samples were collected 2 h post antibiotic exposure and subjected to RNA sequence analysis. Transcriptomic quantification of RNA representing duplicated samples generated highly similar gene expression data. Exposure to sub-inhibitory concentration [0.5 x MIC (minimal inhibitory concentration)] of doxycycline or ciprofloxacin modulated the expression of 237 or 8 genes, respectively, while exposure to an inhibitory concentration (1 x MIC) resulted in the modulation of 583 or 234 genes, respectively. Amongst the genes modulated upon doxycycline exposure upregulation of 31 genes encoding for translation-functions could be distinguished, as well as downregulation of 14 genes encoding for functions involved in DNA transcription and repair. Ciprofloxacin exposure impacted differently the RNA sequence profile of the pathogen, resulting in upregulation of 27 genes encoding mainly DNA replication and repair functions, transmembrane transporters and molecular chaperons. In addition, 15 downregulated genes were involved in translation processes.
作为旨在提出一种替代方法的研究的一部分,通过定量特定标记基因和基因集的表达水平变化来快速确定抗生素敏感性,本研究培养了毒力细菌弗朗西斯菌在存在环丙沙星或强力霉素的抑制/亚抑制浓度的情况下生长,并通过差异表达分析和功能注释阐明了它们的转录组谱。
为了鉴定弗朗西斯菌对环丙沙星或强力霉素暴露的反应中差异表达的基因(DEGs),本研究进行了 RNA 测序,环丙沙星和强力霉素是土拉菌病治疗的首选抗生素。因此,在抗生素暴露后 2 小时收集 RNA 样本并进行 RNA 序列分析。来自重复样本的 RNA 转录组定量生成了高度相似的基因表达数据。亚抑制浓度(0.5xMIC(最小抑制浓度))的强力霉素或环丙沙星暴露分别调节了 237 或 8 个基因的表达,而抑制浓度(1xMIC)暴露分别导致 583 或 234 个基因的表达调节。在强力霉素暴露后调节的基因中,31 个编码翻译功能的基因上调,14 个编码 DNA 转录和修复功能的基因下调。环丙沙星暴露对病原体的 RNA 序列谱产生了不同的影响,导致 27 个编码主要 DNA 复制和修复功能、跨膜转运蛋白和分子伴侣的基因上调,此外,15 个下调基因参与翻译过程。