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. 2014 Aug 14;10(8):e1004316. doi: 10.1371/journal.ppat.1004316

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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(A) Schematic description of the procedure for preparing CfaA/B complex. (B) SDS-PAGE of eluted fractions from Ni-NTA affinity column for wild-type CfaA/B complex and various CfaA mutants detected by Coomassie Brilliant Blue staining. From lane 1 to 14 are CfaB co-purified with CfaA variants: No CfaA, wild type CfaA, K9A, T44A/E45A/E46A, E86A, T112A, L114A, V116A, I118A, Y120A, R125A, R154A, K164-N171(A×8) and C163S/C172S. Unbound CfaA in flow-through fractions were also subjected to SDS-PAGE analysis and detected by immunoblot with specific antibody against CfaB. (C) Ratios between CfaA variant and CfaB were obtained from densitometry analysis averaged over six independent SDS-PAGE experiments shown in (B). Error bars are indicative of standard deviations among independent experiments.