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. 2013 Dec 30;11:122. doi: 10.1186/1741-7007-11-122

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Copyright © 2013 Baumgart et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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IpsA in Corynebacterium diphtheriae and Mycobacterium tuberculosis.Corynebacterium glutamicum ΔipsA carrying the plasmids pAN6, pAN6-DIP1969 or pAN6-rv3573 and C. glutamicum wild-type with pAN6 as control were cultivated in CGXII with glucose without isopropyl β-d-1-thiogalactopyranoside (IPTG) (A) or with 50 μM IPTG (B). (C) Microscopic phenotypes of the complemented strains. DNA was stained with 4’,6-diamidino-2-phenylindole (DAPI) (cyan) and lipophilic regions with nile red (red), scale bar 5 μm. (D) Oligonucleotides (30 bp, 1 μM for DIP0021 and DIP0115, 0.5 μM for Rv0047 and the negative control) covering the predicted binding sites in the promoter regions of the respective genes were incubated with IpsA at the given concentrations and analyzed on 15% native polyacrylamide gels. In M. tuberculosis, Rv0046 is organized in an operon with Rv0047. A binding site was identified within the open reading frame (ORF) of Rv0047, suggesting the occurrence of a second promoter upstream of ino1. (E) IpsA binding motif derived from the high affinity C. glutamicum targets and the C. diphtheriae and M. tuberculosis binding sites.