Fig 5.

DNA binding properties of Rot, SaeR, and Rot/SaeR complexes. (A) EMSA of 2 pmol of purified Rot, 37.5 pmol of purified phospho-SaeR, or a mixture of Rot and phospho-SaeR incubated with 40 fmol of either the ssl7, ssl9, or ssl11 promoter containing a biotin tag. DNA was visualized using streptavidin DyLight. The arrow indicates free probe. (B) Immunoprecipitation of FLAG-tagged Rot from the indicated S. aureus Newman strains. The samples were separated using SDS-PAGE, transferred to nitrocellulose, and immunoblotted (IB) for FLAG to detect Rot or for SaeR (top) or used as temple for PCR amplification of the ssl7 and ssl9 promoters (bottom).