FIGURE 7.

Characterization of bba34 transposon mutant and complemented strains in vitro. (A) Cartoon schematic of wild-type (WT), bba34 transposon mutant (Δa34tn) and allelic-exchange strategy used to generate bba34 complement (bba34comp). Arrows shown above the WT loci indicate location of primers used to amplify upstream and downstream fragments in bba34comp allelic replacement construct. qRT-PCR (B) and immunoblot (C) analyses of WT, Δa34tn and a34comp strains following temperature-shift in vitro. Transcript levels for bba34 were normalized using a TaqMan-assay for flaB. Error bars indicate standard errors of the mean (3 biological replicates per strain, assayed in quadruplicate). p-Values for pairwise comparisons were determined using a two-tailed t-test. Whole cell lysates were separated on a 12.5% SDS polyacrylamide gel and then either stained with Coomassie blue or transferred to nitrocellulose for immunoblotting with polyclonal antisera against strain B31 OspC (generated as part of these studies), RpoS (Hyde et al., 2007) and FlaB (Caimano et al., 2005). Molecular markers (kDa) are shown on the left.