Borrelia peptidoglycan interacting Protein (BpiP) contributes to the fitness of Borrelia burgdorferi against host-derived factors and influences virulence in mouse models of Lyme disease
Fig 5
Generation of the bpiP mutant and complement strains.
(A) Schematic representation of the genetic manipulation performed to generate bpiP (bb_0167) mutant and cis-complemented strains. (B) Total RNA from wt, mt and ct grown at pH7.6/32°C was converted to cDNA and quantitative real-time PCR was performed as described in the Materials and Methods section. The levels of bpiP transcripts were absent as expected in the mt compared the control strains. The change in the Ct value for each sample was subjected to an unpaired t test in GraphPad Prism software. The asterisk indicates the levels of significance as follows: *** p<0.001. Total protein lysates from wt (Lane 1), mt (Lane 2) and ct (Lane 3) were separated on 12% SDS-PAGE gel and (C) stained with Coomassie blue or (D) transferred to PVDF membrane for immunoblot analysis using antibodies specific to streptomycin resistance (α-Strep, mt) or gentamicin resistance (α-Gent, ct) proteins synthesized due to the expression of aadA and gentR resistance genes under the control of borrelial promoter PflgB.