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. 2017 May 11;8(8):623–627. doi: 10.1007/s13238-017-0417-3

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© The Author(s) 2017

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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.

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Anti-ciprofloxacin complementation assay. (A) A serial dilution of cell culture spotted onto solid medium containing 100 µg/mL ampicillin and 5 ng/mL ciprofloxacin. Compared with the empty-vector (pET21a) and predicted protonation-site mutant D407N containing strain, the wild type AcrB constructed in pET21a showed apparent resistance against ciprofloxacin. Leaky expression of AcrB was confirmed using anti-His immunoblotting. (B) Mutational effects in the two signaling motif regions. (C) Mutational effects in the amphipathic helix α6-7. 12-GS is the variant that the entire α6-7 was replaced with 12 Gly-Ser repeats. (D) Mutational effects in the region of titratable key residues