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. 2017 Nov 27;7:16395. doi: 10.1038/s41598-017-16625-6

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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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Aacec B peptide binds to the putative binding motif TTGG(A/C)A present in four DNA fragments of AaPPO 3. (a) A schematic representation of the AaPPO 3 gene. Open boxes with roman numerals indicate exons and the nucleotide length of introns and exons are indicated by numbers. The green boxes indicate the copper binding site Cu A and Cu B. (b–f) DNA pull-down assay showing Aacec B peptide binding to AaPPO 3 DNA. The upper figures show schematic representations of the AaPPO 3 DNA fragments. The nucleotide lengths are indicated by numbers. The lower figures are the PCR analyses of the AaPPO 3 DNA fragments that are able to bind to Aacec B-conjugated beads. The arrows point to the PCR amplifications of the AaPPO 3 DNA fragments binding to the Aacec B-conjugated beads. (g) DNA sequence logo representing the TTGG(A or C)A binding motif that was identified from the four AaPPO 3 DNA fragments (5′ end-1-2, Cu A1, Cu B2 and Cu B3) using the MEME suite. The height of each letter represents the relative frequency of occurrence of the nucleotide at each position. (h) The nucleotide sequences of the four AaPPO 3 DNA fragments (5′ end-1-2, Cu A1, Cu B2 and Cu B3) containing the TTGGAA and TTGGCA putative motifs (indicated in red). Uncropped images are shown in Supplementary Figure S5.