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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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Analyses of Riemerella anatipestifer AS87_RS09170 gene and deduced amino acid sequence. (A) Polymerase chain reaction (PCR) analysis. The AS87_RS09170 gene was amplified from all R. anatipestifer strains tested. Lane M, DL2000 DNA Marker (Takara); lanes 1–5: R. anatipestifer serotype 1 strains GTB1, GTB2, CQ1, CQ3 and NJ4; lanes 6–10, R. anatipestifer serotype 2 strains GDO-3, JY1, JY2, NJ3 and SC2; lanes 11–15, R. anatipestifer serotype 10 strains GDO-1, HXb2, YXb1, YXb11 and YXL1; lanes 16–20, R. anatipestifer serotype 15 strains SQ007, SQ003, SQ004, YGT002 and MA001; lanes 21–25, R. anatipestifer undefined serotype strains GDO-6, GDO-7, G46, G77 and JY-6; lane 26, negative control of Yb2ΔbioF; lane 27, positive control of Yb2. A 1158-bp fragment was amplified from all 25 R. anatipestifer strains tested. The full-length gel is presented in Supplementary Figure S2. (B) Identity and divergence analysis. The predicted R. anatipestifer 8-amino-7-oxononanoate synthase (AONS) and other known AONS were compared. The crystallized AONS sequences [Paraburkholderia xenovorans (pdb5JAY), Francisella tularensis (pdb4IW7), Escherichia coli (pdb1DJE), Burkholderia multivorans (pdb5VNX) and Mycobacterium smegmatis (pdb3WY7)] were retrieved from the Uniprot database and later aligned with Clustal W algorithm in the MegAlign program from the DNASTAR Lasergene suite. (C) Multiple sequence alignment. Strictly conserved residues in all sequences are boxed in black. Residues forming the conserved pyridoxal 5′-phosphate binding pocket are indicated by a black inverted triangle (G97, Y98, N101, E168, D197, H200, T228 and K231).