ArticleProtein expression and purification2017
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis.
Article in Protein expression and purification, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 9 citations in OpenAlex.
- Single-Molecule Methods to Investigate Mechanisms of Transcription by RNA Polymerase ofbioRxiv : the preprint server for biology · 2026Article
- Article
- Pleomorphic effects of three small-molecule inhibitors on transcription elongation bybioRxiv : the preprint server for biology · 2025Article
- Structural insights into the transcription activation mechanism of the global regulator GlnR from actinobacteria.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- DNA structural properties of DNA binding sites for 21 transcription factors in the mycobacterial genome.Frontiers in cellular and infection microbiology · 2023Article
- Mycobacterium tuberculosis CarD, an essential global transcriptional regulator forms amyloid-like fibrils.Scientific reports · 2018Article
- Association of ω with the C-Terminal Region of the β' Subunit Is Essential for Assembly of RNA Polymerase in Mycobacterium tuberculosis.Journal of bacteriology · 2018Article
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Authors and funding
6 authors at 3 institutions in 2 countries.
Funding
Abstract
Recent publications have shown that active RNA polymerase (RNAP) from Mycobacterium tuberculosis (MtbRNAP) can be produced by expressing all four subunits in a single recombinant Escherichia coli strain [1-3]. By reducing the number of plasmids and changing the codon usage of the Mtb genes in the co-expression system published by Banerjee et al. [1], we present a simplified, detailed and reproducible protocol for the purification of recombinant MtbRNAP containing the ω subunit. Moreover, we describe the formation of ternary elongation complexes (TECs) with a short fluorescence-labeled RNA primer and DNA oligonucleotides, suitable for transcription elongation studies. The purification of milligram quantities of the pure and highly active holoenzyme omits ammonium sulfate or polyethylene imine precipitation steps [4] and requires only 5 g of wet cells. Our results indicate that subunit assemblies other than α
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