ArticleNucleic acids research2015
Mycobacterial RNA isolation optimized for non-coding RNA: high fidelity isolation of 5S rRNA from Mycobacterium bovis BCG reveals novel post-transcriptional processing and a complete spectrum of modified ribonucleosides.
Article in Nucleic acids research, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 18 citations in OpenAlex.
- Novel molecular assessment method using precursor ribosomal RNA to rapidly quantify hard-to-monitor pathogens in aqueous suspensions: application to ultraviolet disinfection ofJournal of water and health · 2025Article
- Are Bacterial Processes Dependent on Global Ribosome Pausing Affected by tRNA Modification Defects?Journal of molecular biology · 2025Review
- Molecular Coping Mechanisms: Reprogramming tRNAs To Regulate Codon-Biased Translation of Stress Response Proteins.Accounts of chemical research · 2023Article
- An RNA modification enzyme directly senses reactive oxygen species for translational regulation in Enterococcus faecalis.Nature communications · 2023Article
- Mycobacterium tuberculosis whiB3 and Lipid Metabolism Genes Are Regulated by Host Induced Oxidative StressMicroorganisms · 2022Article
- Quantitative mapping of the cellular small RNA landscape with AQRNA-seq.Nature biotechnology · 2021Article
- Comparing mRNA expression and protein abundance in MDRBiotechnology reports (Amsterdam, Netherlands) · 2021Article
- Cascade of deoxyribozymes for the colorimetric analysis of drug resistance in Mycobacterium tuberculosis.Biosensors & bioelectronics · 2020Article
- The response of Synechococcus sp. PCC 7002 to micro-/nano polyethylene particles - Investigation of a key anthropogenic stressor.PloS one · 2020Article
- tRNA-mediated codon-biased translation in mycobacterial hypoxic persistence.Nature communications · 2016Article
- A Platform for Discovery and Quantification of Modified Ribonucleosides in RNA: Application to Stress-Induced Reprogramming of tRNA Modifications.Methods in enzymology · 2015Article
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Authors and funding
6 authors at 4 institutions in 2 countries.
Funding
Abstract
A major challenge in the study of mycobacterial RNA biology is the lack of a comprehensive RNA isolation method that overcomes the unusual cell wall to faithfully yield the full spectrum of non-coding RNA (ncRNA) species. Here, we describe a simple and robust procedure optimized for the isolation of total ncRNA, including 5S, 16S and 23S ribosomal RNA (rRNA) and tRNA, from mycobacteria, using Mycobacterium bovis BCG to illustrate the method. Based on a combination of mechanical disruption and liquid and solid-phase technologies, the method produces all major species of ncRNA in high yield and with high integrity, enabling direct chemical and sequence analysis of the ncRNA species. The reproducibility of the method with BCG was evident in bioanalyzer electrophoretic analysis of isolated RNA, which revealed quantitatively significant differences in the ncRNA profiles of exponentially growing and non-replicating hypoxic bacilli. The method also overcame an historical inconsistency in 5S rRNA isolation, with direct sequencing revealing a novel post-transcriptional processing of 5S rRNA to its functional form and with chemical analysis revealing seven post-transcriptional ribonucleoside modifications in the 5S rRNA. This optimized RNA isolation procedure thus provides a means to more rigorously explore the biology of ncRNA species in mycobacteria.
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