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. 2015 Mar 12;6:170. doi: 10.3389/fmicb.2015.00170

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Copyright © 2015 Rojas Murcia, Lee, Waridel, Maspoli, Imker, Chai, Walsh and Reimmann.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of hexahistidine-tagged proteins purified by Ni-NTA chromatography. (A) 6% Separating gel with protein ladder (lane 1), and AmbE (231.5 KDa, lane 2) and AmbB (137 KDa, lane 3) purified from BL21[DE3]/pLys carrying pME9714 and pME9713, respectively. (B) 10% Separating gel with protein ladder (lane 1), and AmbC (41.5 KDa, lane 2), and AmbD (39.5 KDa, lane 3) purified from BL21[DE3] carrying pME9718 and pME9717, respectively. (C) 6% Separating gels of purified mutant proteins. Lanes 1 and 3, protein ladders; lane 2, AmbB_S768A expressed from pME10327; lane 4, AmbE_{D644A,K1230T,S1958A} expressed from pME10330; lane 5, AmbE_S1958A expressed from pME10328; lane 6, AmbE_S1819A expressed from pME10317; lane 7, AmbE_S1286A expressed from pME10337. All proteins were purified from BL21[DE3]/pLys carrying the respective plasmid.