Objectives: The non-coding RNA rprA can increase the resistance to ampicillin in Escherichia coli.
Methods: 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.
Results: 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).
Conclusion: 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.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.micpath.2019.02.021 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Aptazyme-directed A-to-I RNA editing.
Methods Enzymol
January 2025
Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Science, Chengdu, P.R. China. Electronic address:
As a promising therapeutic approach, the RNA editing process can correct pathogenic mutations and is reversible and tunable, without permanently altering the genome. RNA editing mediated by human ADAR proteins offers unique advantages, including high specificity and low immunogenicity. Compared to CRISPR-based gene editing techniques, RNA editing events are temporary, which can reduce the risk of long-term unintended side effects, making off-target edits less concerning than DNA-targeting methods.
View Article and Find Full Text PDFRestoration of G to A mutated transcripts using the MS2-ADAR1 system.
Methods Enzymol
January 2025
Area of Bioscience and Biotechnology, School of Materials Science, Japan Advanced Institute of Science and Technology, Asahidai, Nomicity, Ishikawa, Japan. Electronic address:
Site-directed RNA editing (SDRE) holds significant promise for treating genetic disorders resulting from point mutations. Gene therapy, for common genetic illnesses is becoming more popular and, although viable treatments for genetic disorders are scarce, stop codon mutation-related conditions may benefit from gene editing. Effective SDRE generally depends on introducing many guideRNA molecules relative to the target gene; however, large ratios cannot be achieved in the context of gene therapy applications.
View Article and Find Full Text PDFEn masse evaluation of RNA guides (EMERGe) for ADARs.
Methods Enzymol
January 2025
Department of Chemistry, University of California, Davis, 1 Shields Ave, Davis, CA, United States. Electronic address:
Adenosine Deaminases Acting on RNA (ADARs) convert adenosine to inosine in duplex RNA, and through the delivery of guide RNAs, can be directed to edit specific adenosine sites. As ADARs are endogenously expressed in humans, their editing capacities hold therapeutic potential and allow us to target disease-relevant sequences in RNA through the rationale design of guide RNAs. However, current design principles are not suitable for difficult-to-edit target sites, posing challenges to unlocking the full therapeutic potential of this approach.
View Article and Find Full Text PDFLeveraging Saccharomyces cerevisiae for ADAR research: From high-yield purification to high-throughput screening and therapeutic applications.
Methods Enzymol
January 2025
Life Science, Bar Ilan University, Ramat Gan, Israel. Electronic address:
Saccharomyces cerevisiae, a model eukaryotic organism with a rich history in research and industry, has become a pivotal tool for studying Adenosine Deaminase Acting on RNA (ADAR) enzymes despite lacking these enzymes endogenously. This chapter reviews the diverse methodologies harnessed using yeast to elucidate ADAR structure and function, emphasizing its role in advancing our understanding of RNA editing. Initially, Saccharomyces cerevisiae was instrumental in the high-yield purification of ADARs, addressing challenges associated with enzyme stability and activity in other systems.
View Article and Find Full Text PDFStructural analysis of human ADAR2-RNA complexes by X-ray crystallography.
Methods Enzymol
January 2025
Department of Chemistry, University of California, Davis, CA, United States; Department of Molecular and Cellular Biology, University of California, Davis, CA, United States. Electronic address:
Adenosine deaminases acting on RNAs (ADARs) are a class of RNA editing enzymes found in metazoa that catalyze the hydrolytic deamination of adenosine to inosine in duplexed RNA. Inosine is a nucleotide that can base pair with cytidine, therefore, inosine is interpreted by cellular processes as guanosine. ADARs are functionally important in RNA recoding events, RNA structure modulation, innate immunity, and can be harnessed for therapeutically-driven base editing to treat genetic disorders.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!