FIG. 6.

Analysis of the composition of T-pilus fractions. Strains C58, CB1002 (ΔvirB2), CB1005 (ΔvirB5), and CB1008 (ΔvirB8) and complemented variants were cultivated on AB minimal medium in the absence (−) of AS (lane 1) or in the presence (+) of AS (lanes 2 to 7) for virulence gene induction, followed by shearing of cells and ultracentrifugation for separation of extracellular high-molecular-mass structures (ultracentrifugation pellet) and low-molecular-mass proteins released from the cells (ultracentrifugation supernatant). Samples were separated by SDS-PAGE, followed by Western blotting with VirB2-, VirB5-, and Tzs-specific antiserum. Lanes: 1, C58 without AS; 2, C58 with AS; 3, CB1002; 4, CB1005; 5, CB1002 pTrcB2; 6, CB1005 pTrcB5; 7, CB1008. The arrowhead points to the reduced amount of Tzs in the ultracentrifugation pellet from CB1005, and molecular masses of reference proteins are shown on the right (in kilodaltons).