Nour Danesh Institute of Higher Education, Department of Biology, Isfahan, Iran.
Department of Genetics, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
Microb Pathog. 2019 Apr;129:266-270. doi: 10.1016/j.micpath.2019.02.021. Epub 2019 Feb 22.
The non-coding RNA rprA can increase the resistance to ampicillin in Escherichia coli.
Bacterial DNA was extracted by boiling method and then amplified using polymerase chain reaction (PCR) with two different primer sets. Recombinant pET28a/rprA-sense and -antisense plasmids were separately transferred into the competent E. coli BL21 (DE3) by chemical methods using heat shock. The expression was analyzed at the RNA level using Semi quantitative RT PCR. The turbidity difference between the bacteria was checked by Broth Dilution method.
The statistical analysis showed that the turbidity difference between the up regulated and control bacteria is significant (p value < 0.0001). The ANOVA test also showed the significant difference between the down regulated and control bacteria (p value < 0.0001).
Considering this mechanism, there are some reports indicating the role of rprA in antibiotic resistance. However, the role of rprA in ampicillin resistance is remained to be unknown. The aim of this study was to analyze the up regulation and down regulation of rprA and check their effects on ampicillin resistance in Escherichia coli. It was found that the up regulation and down regulation of rprA can lead into more antibiotics resistance and susceptibility, respectively. Our results showed the potential role of rprA expression in the response to ampicillin stress in E. coli.
非编码 RNA rprA 可以提高大肠杆菌对氨苄青霉素的抗性。
采用煮沸法提取细菌 DNA，然后使用聚合酶链反应（PCR）用两种不同的引物对进行扩增。分别通过化学方法（热激）将重组 pET28a/rprA- sense 和 -antisense 质粒转入感受态大肠杆菌 BL21（DE3）中。采用半定量 RT-PCR 分析 RNA 水平的表达。通过肉汤稀释法检查细菌浊度差异。
统计分析显示，上调组和对照组细菌的浊度差异具有统计学意义（p 值<0.0001）。方差分析也显示下调组和对照组细菌之间存在显著差异（p 值<0.0001）。
考虑到这种机制，有一些报道表明 rprA 在抗生素抗性中起作用。然而，rprA 在氨苄青霉素抗性中的作用尚不清楚。本研究的目的是分析 rprA 的上调和下调及其对大肠杆菌氨苄青霉素抗性的影响。结果发现 rprA 的上调和下调分别导致了更高的抗生素抗性和敏感性。我们的结果表明 rprA 表达在大肠杆菌对氨苄青霉素应激的反应中具有潜在作用。
