FIG. 1.
Surface expression of Sgp1 and Sgp2 in wild-type strain 89/1591. (A) Genetic organization of the srtG pilus cluster. This schema is constructed on the basis of the draft genome sequence of S. suis 89/1591, updated in February 2009 (GenBank accession no. AAFA03000013). Gray arrows, genes encoding CWSS-containing proteins (sgp1 and sgp2); black arrow, sortase-encoding gene (srtG); open arrows outlined in gray, genes flanking the cluster; small arrowheads, annealing positions of the primers used to generate His-tagged recombinant Sgp1 and Sgp2. (B) Immunoblots with anti-Sgp1 (α-Sgp1; left panel) and anti-Sgp2 (α-Sgp2; right panel) rabbit pAbs using bacterial cells at different growth temperatures (37°C, 33°C, 28°C, or 23°C). Bacteria were grown in THB until the optical density at 600 nm reached 0.35, which corresponds to the mid-log phase. Each lane contains cell wall fractions extracted from cells in 750-μl cultures with an optical density at 600 nm of 0.35. The calculated molecular weights of Sgp1m and Sgp2m are 52,045 and 104,403, respectively. Numbers to the left of the gels are molecular weights (in thousands).