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. 2018 Feb 7;10(2):74. doi: 10.3390/toxins10020074

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Location of the yonT/sr6/yoyJ/yonU locus and determination of transcription start sites. (A) Schematic representation of the location of the yonT/sr6/yoyJ/yonU locus on the B. subtilis chromosome. The directions of transcription are indicated by arrows; (B) Mapping of the 5′ ends of yonT mRNA, yonU mRNA and SR6. Sanger sequencing reactions of heterologous genes were used as references to calculate the location of the transcription start signals. Outer right lanes show primer extension reactions on total RNA of B. subtilis DB104 with 5′-labelled primer SB2648 (sr6), SB2646 (yonT), and SB2647 (yonU) loaded in parallel to the sequencing reactions (transcription start sites are indicated by arrows). Below: sr6, yonT and yonU sequences around the TSS are shown. Dots symbolize the mapped TSS; (C) Sequence of the yonT/yoyJ/sr6 locus. −10 boxes of the yonT and the sr6 promoters are underlined. Start and direction of transcription are indicated by arrows and transcription termination is indicated by ‘term’. The yonT/SR6/yoyJ complementary region is highlighted in light grey. Start and stop codons of the yonT and yoyJ ORFs are in bold and the putative SD sequences are boxed.