Fig. 1.

Raffinose transport system. (A) The raf locus in S. pneumoniae D39. The large boxed arrows represent ORFs. Below, the corresponding gene numbers are listed as well as the size of the gene in base pairs (bp). Small arrows indicate putative promoter sites and orientation of corresponding transcripts. CRE, catabolite response element. (B) The rafK locus in S. pneumoniae D39. The rafK gene has its own putative promoter with a predicted ribosomal binding site, and a predicted terminator is located immediately after the gene. (C) Western blot of bacterial lysates using anti-RafK rabbit polyclonal antibodies (lanes 1 to 6), rabbit preimmune serum (lane 7), or anti-lipoic acid rabbit polyclonal antibodies (lane 8). Lanes: 1, D39 lysate (parental wild-type); 2, D39-ΔRafK-Janus lysate (RafK Janus mutant); 3, D39-ΔRafK lysate (RafK deletion mutant); 4, D39-rRafK lysate (allelic repair of RafK Janus mutant with full-length RafK); 5, D39-rRafKΔlipErm lysate (allelic repair of RafK Janus mutant with RafK lacking the lipoyl domain); 6, recombinantly expressed RafK protein; 7, D39 lysate probed with preimmune serum; 8, D39 lysate lysate probed with rabbit anti-lipoic acid antibodies. The RafK-specific band is indicated in the figure with an arrow. The anti-RafK polyclonal serum also contained unspecific reactivity against a 50-kDa protein that was present also in preimmune sera (lane 7) and could not be adsorbed away.