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. 2017 Aug 30;7:9982. doi: 10.1038/s41598-017-10155-x

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© The Author(s) 2017

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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cd2068 C. difficile mutant exhibited increased drug susceptibility. (a) Verification of the cd2068 mutant in C. difficile. (Top panel) schematic cartoons illustrated the cd2068 gene before and after retro-transposition of the intron, arrows indicate primer used for verification. (Bottom panel) Agarose gel electrophoresis of PCR products amplified using corresponding set of primers. M represents DNA ladder; Lane 1 represents pMTL007C-E2:cd2068 plasmid; Lanes 2, 4 and 6 represent C. difficile Δerm (control); Lanes 3, 5 and 7 represent cd2068 mutant. Lanes 1–3, 4–5, and 6–7 were amplified using RAM-F/RAM-R, EBS universal/CD2068-R, and CD2068-F/CD2068-R, respectively. (b) Disruption of cd2068 in C. difficile enhanced antibiotic susceptibility, which was genetically complemented with the cd2068 gene. Agar well diffusion assays were performed on the cd2068 mutant versus control C. difficile as well as the cd2068 mutants carrying pMTL84151 and pMTL-cd2068 in the presence of various concentrations of ciprofloxacin.