MsvR binding and regulatory targets assessed by EMSA. (a) Sequences of the 50 bp region of Ma PmsvR used to confirm MaMsvR binding to boxes A and B. Sequence changes within the binding boxes are shown. (b) EMSA assays with the template (50 nM) variations shown in (a) and 1 μM (20-fold excess over DNA) reduced MaMsvR (R, 5 mM DTT). A 50 bp region of Ma PmsvR was included as a binding control. The gel wells are indicated (W). (c) EMSA analysis with reduced MaMsvR (R, 5 mM DTT) and its own promoter (Ma PmsvR, 10 nM), various intergenic regions of an oxidative stress response cluster (Ma P4664, P3734, P3736, 10 nM) as well as the control Ma histone A promoter (Ma PhmaA, 10 nM). A region of rpoK (10 nM) was tested for binding because an MsvR binding site (TTCGN8CGAA) is present in the coding region. The gel wells are indicated (W).
Isom et al. BMC Microbiology 2013 13:163 doi:10.1186/1471-2180-13-163