Figure 4.

In vitro characterization of the specific binding of BsSpo0J to parS DNA duplexes. EMSA of wild-type BsSpo0J and mutants binding to either a radioactively labeled 39-bp parS DNA substrate supplemented with cold 39-bp scrambled parS competitor DNA as shown in (A, C and E), or a labeled 24-bp parS substrate without competitor DNA as shown in (B, D and F) (see ‘Materials and Methods’ section). Protein concentrations were 0.2, 0.4, 0.8 μM in (A and B) and 0.2, 0.4, 0.8 and 1.0 μM in (C–F). Asterisk and arrow indicate position of the wells and free DNA respectively in each gel. See Supplementary Figures S13 and 14 for results of all characterized mutants.