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. 2018 Jul 13;8:10652. doi: 10.1038/s41598-018-28843-7

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Functional analysis of the IR Rho-independent transcription terminator. (a) Design of reporter constructs. To assess the functionality of the putative IR transcriptional terminator TERM 264, the constitutive B. thailandensis ribosomal gene 12 promoter (P_S12) was inserted upstream of the penA’-lacZ transcriptional fusion such that P_S12 was centered either 134 bp or 24 bp upstream of TERM 264 (indicated by the lollipop) or 134 bp or 24 bp upstream of a clean TERM 264 deletion (indicated by Δ). The center of P_S12 promoter sequence is marked with a filled triangle in the shown sequence. These fragments were cloned into a mini-Tn7-lacZ transcriptional fusion vector and integrated into the Bp82.27 chromosome. (b) β-Gal activity in strains harboring single-copy lacZ reporter constructs. The strains resulting from chromosomal integration of P_S12-penA’-lacZ fusions were Bp82.374 (P_S12 134 bp upstream of TERM 264), Bp82.380 (P_S12 24 bp upstream of TERM 264), Bp82.375 (P_S12 134 bp upstream of ΔTERM 264) and Bp82.381 (P_S12 24 bp upstream of ΔTERM 264). Bp82.363 served as empty vector control. β-Gal activities were measured in triplicate on three separate days and are expressed in Miller units. Error bars indicate standard deviation from the mean.