Figure 1. Genotypes and phenotypes of p66 strains used in this study.

(A) Schematic representation of p66 loci of wild-type (WT), mutant (Δp66), and complemented (p66^cc and p66^cp) B. burgdorferi. The scale indicates the size and spacing of the genes in base pair units. GentR refers to the gentamicin resistance cassette, aacC1 from Tn1696, driven by the flgB promoter (Elias et al., 2003). KanaR refers to the kanamycin resistance cassette, aphI from Tn903, driven by the flaB promoter (Bono et al., 2000). (B) Western blot of whole-cell lysates from in vitro grown B. burgdorferi of indicated strains. Two clones of p66^cc are shown to represent a clone with each orientation of the gentamicin resistance cassette with respect to p66. Lysates were run on an SDS-PAGE gel and transferred to an Immobilon membrane that was probed with antibodies detecting P66 and flagellin (as a loading control). Colorimetric methods were used for visualization. The values on the left represent molecular size in kilodaltons. (C) Quantification by densitometry of a parallel blot of that shown in panel B with chemiluminescence used for detection. Panels B and C represent one of three independent clones with similar phenotypes. P66 expression was normalized to flagellin expression.