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. 2006 Jan;74(1):108–117. doi: 10.1128/IAI.74.1.108-117.2006

FIG. 6.

FIG. 6.

Expression of the B. suis virB1+3-12 operon induces periplasmic leakage in A. tumefaciens. Cells were cultivated on AB minimal medium plates at 20°C for 3 days, followed by shearing of the cells and analysis of protein content of subcellular fractions after SDS-PAGE and Western blotting: cell lysate (C), ultracentrifugation supernatant (S), and pellet (P [pilus fraction]). (A) A. tumefaciens wild-type C58 grown under noninducing (−AS) or virulence gene-inducing conditions (+AS). (B) A. tumefaciens wild type grown under noninducing (lanes 1) or virulence gene-inducing conditions (lanes 2) and Ti plasmid-free strain UIA143 grown without IPTG (lanes 3) or with IPTG (lanes 4). Strain UIA143 was grown with different virB operon plasmids in the presence of IPTG as follows: lanes 5, pTrc300; lanes 6, pTrcB3-12; lanes 7, pTrcB1+3-12; and lanes 8, pTrcB2-12. An arrowhead indicates periplasmic protein AcvB released into the supernatant by shearing. Antisera detected A. tumefaciens VirBa proteins or B. suis VirB proteins. The molecular masses of reference proteins are shown on the right.