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Viruses 2018, 10(11), 595; https://doi.org/10.3390/v10110595

Comparative Genomics and Characterization of the Late Promoter pR’ from Shiga Toxin Prophages in Escherichia coli

Department of Agriculture, Food and Nutritional Science, University of Alberta, Edmonton, AB T6G 2P5, Canada
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Received: 29 September 2018 / Revised: 26 October 2018 / Accepted: 27 October 2018 / Published: 31 October 2018
(This article belongs to the Special Issue Viruses of Microbes V: Biodiversity and Future Applications)
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Abstract

Shiga-toxin producing Escherichia coli (STEC) causes human illness ranging from mild diarrhea to death. The bacteriophage encoded stx genes are located in the late transcription region, downstream of the antiterminator Q. The transcription of the stx genes is directly under the control of the late promoter pR’, thus the sequence diversity of the region between Q and stx, here termed the pR’ region, may affect Stx toxin production. Here, we compared the gene structure of the pR’ region and the stx subtypes of nineteen STECs. The sequence alignment and phylogenetic analysis suggested that the pR’ region tends to be more heterogeneous than the promoter itself, even if the prophages harbor the same stx subtype. Furthermore, we established and validated transcriptional fusions of the pR’ region to the DsRed reporter gene using mitomycin C (MMC) induction. Finally, these constructs were transformed into native and non-native strains and examined with flow cytometry. The results showed that induction levels changed when pR’ regions were placed under different regulatory systems. Moreover, not every stx gene could be induced in its native host bacteria. In addition to the functional genes, the diversity of the pR’ region plays an important role in determining the level of toxin induction. View Full-Text
Keywords: Shiga toxin prophage; genomic characterization; flow cytometry; microscope; phage induction efficiency; sequence diversity Shiga toxin prophage; genomic characterization; flow cytometry; microscope; phage induction efficiency; sequence diversity
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Zhang, L.X.; Simpson, D.J.; McMullen, L.M.; Gänzle, M.G. Comparative Genomics and Characterization of the Late Promoter pR’ from Shiga Toxin Prophages in Escherichia coli. Viruses 2018, 10, 595.

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