Figure 2. ParB binds preferentially to parS in vitro.

(A) Agarose EMSA on after incubating different amounts of ParB with a ³²P-labeled 3120 bp DNA fragment containing the parS-cluster, (B) or with a 3005 bp DNA containing the Mx8 phage attP sequence. (C) Agarose EMSA of the binding of ³²P-labeled parS-cluster to ParB, or to ParB previously incubated with higher amounts of unlabeled attP DNA fragment, as indicated. (D) Agarose EMSA of the binding of ³²P-labeled attP fragment to ParB, or to ParB previously incubated with higher amounts of unlabeled parS-cluster, as indicated. (E) EMSA using a 6% polyacrylamide gel after incubating a ³²P-labeled 50 bp DNA fragment containing the M. xanthus parS consensus sequence with increasing amounts of ParB, as indicated (F) or with this ³²P-labeled 50 bp DNA fragment containing 11 point mutations of the most conserved base pairs of parS.