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. 2017 Oct 16;8:2013. doi: 10.3389/fmicb.2017.02013

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Copyright © 2017 Zhang, Wu, Zhu, Liu, Wang, Cao, Chen, Tao and Pang.

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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Electrophoretic mobility shift assays (EMSA) and transcriptional analysis of genes with upstream sequences similar to the MtrA_MTB recognition sequence. (A,B) EMSA analyses with MtrA_SCO and 59-bp oligonucleotides containing sequences similar to the MtrA_MTB recognition sequence. A fixed amount of labeled oligonucleotide was incubated in reactions containing no MtrA (lane 1), or 1.6 or 3.2, or 4.8 μg MtrA (lanes 2, 3, and 4, respectively). (A) Analysis with sequences upstream of bldD and whiI that were protected by MtrA_SCO in DNase I footprinting (see Figure 5). (B) Analysis with sequences identified in a genome-wide search for matches to the MtrA_MTB consensus recognition sequence. The sequences were located upstream of the named genes. (C) Transcriptional analysis of potential MtrA_SCO target genes in the ΔmtrA mutant. M145 and ΔmtrA were grown on YBP solid medium, and RNA samples were isolated at the indicated times. Expression of hrdB, encoding the major sigma factor in M145, was used as an internal control. The y-axis shows the fold change in expression level in M145 (gray bars) over the level of ΔmtrA (light bars) at each time point, which was arbitrarily set to one. Results are the means (± SD) of triplet experiments.