FIG. 3.
Competitive EMSA with C. trachomatis dnaK CIRCE probes and purified recombinant HrcA. (A) Representation of DNA fragments used in EMSA. CIRCE and promoter elements are marked. (B) Competition with CIRCE-containing restriction fragments. Lane 1, probe alone; lane 2, no competitor; lanes 3 to 7, molar excess of specific competitor relative to labeled probe (2-, 4-, 8-, 16-, and 32-fold); lanes 8 to 12, molar excess of nonspecific competitor (2-, 4-, 8-, 16-, and 32-fold). (C) Competition with CIRCE-containing oligonucleotides. Lane 1, probe alone; lane 2, no competitor; lanes 3 to 6, molar excess of specific competitor (4-, 8-, 16-, and 32-fold); lanes 7 to 10, molar excess of nonspecific competitor (4-, 8-, 16-, and 32-fold). All reactions, with the exception of reactions lacking HrcA, were performed with 0.5 nM labeled probe and 160 nM HrcA.