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. 2018 Oct 2;8:14645. doi: 10.1038/s41598-018-32905-1

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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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Characterization of mutant strain Yb2ΔbioF. (A) Growth curves of Yb2, Yb2ΔbioF and cYb2ΔbioF in tryptic soy broth (TSB). (B) Growth curves of Yb2ΔbioF in TSB, TSB-biotin and TSB-biotin+biotin. Each point represents the mean ± standard for triplicate samples. (*p < 0.05; ***p < 0.001). (C) Immunoblot analysis of the biotinylated protein. Whole-cell proteins of each strain were separated on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel, transferred to nitrocellulose membrane, blotted with streptavidin peroxidase, and visualized by chemiluminescence. Riemerella anatipestifer GroEL was used as a protein loading control. The full-length blots are presented in Supplementary Figure S3. (D) Percoll density gradient centrifugation of the R. anatipestifer strains. The gradient is visualized using fuchsine-stained layers in the marker (M). Lane 1, the wild-type strain Yb2; lane 2, the mutant strain Yb2ΔbioF; lane 3, the complemented strain cYb2ΔbioF.