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. 2020 Jun 25;48(14):7786–7800. doi: 10.1093/nar/gkaa533

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 5. — AusR binds to promoters containing the predicted operator sequence. (A) Sequence logo of the conserved DNA motif detected upstream genes from the AUS. Individual sequences are shown in Supplementary Figure S4. (B) SDS-PAGE analysis of the purified recombinant His-tagged AusR protein. Lane L, molecular weight ladder. (C, D) Electrophoretic mobility shift assays of DNA fragments covering the promoters PausR (C) and PalyA3 (D) with increasing amount of purified His-tagged AusR (from 0 to 38 pmol). (E, F) Alginate trisaccharides prevent AusR binding to its promoters. EMSAs were performed with 20 pmol of purified His-tagged AusR and 60 fmol of Cy5-labeled PausR (E) or PalyA3 (F), in the absence of any ligand or with 100 nmol D-glucose, D-gluconate, ΔGG, ΔMM, ΔMG, D-mannuronate or L-guluronate. The first lane without AusR shows the mobility of free DNA. Δ: 4-deoxy-L-erythro-hex-4-enepyranosyluronate; M: D-mannuronate; G: L-guluronate.