FIG. 2.

Interaction of His-YmoA with E. coli H-NS (A and B) and Y. enterocolitica H-NS (C and D). (A) Coomassie blue-stained SDS-PAGE gel of fractions eluted with increasing imidazole (75, 100, 200, and 200 mM) from Ni²⁺-NTA agarose resin bound to a cleared cellular extract from IPTG-induced E. coli BL21(DE3)(pLysE, pETHISYMOA). (B) Western blotting with H-NS-specific antibodies of fraction 4 from panel A. (C) Lane 1, fraction eluted with 200 mM imidazole from Ni²⁺-NTA-agarose resin containing bound His-YmoA protein. Interacting proteins (H-NS from E. coli) had been previously removed by repeated washing with buffer A-1.0 M KCl. Lanes 2 to 5, fractions eluted with buffer A with increasing concentrations of imidazole (50, 75, 100, and 200 mM) from Ni²⁺-NTA-agarose resin containing bound His-YmoA protein mixed with a cellular extract from Y. enterocolitica Y754. Lane 6 shows unspecific binding of an unknown protein (a) of Y. enterocolitica to the Ni²⁺-NTA-agarose resin. (D) Western blotting with E. coli H-NS-specific antibodies of fractions 5 and 1 from panel C (lanes 1 and 2, respectively). M, molecular mass standard (A and C) or prestained molecular mass standard (B and D).