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. 2013 Jul 3;8(7):e68799. doi: 10.1371/journal.pone.0068799

Figure 2. The ospC operator and mutagenesis strategy.

Figure 2

(A) ospC operator mutations are linked to the kanamycin resistance cassette (flgBp-aphI). The sequence upstream of the ospC gene in B. burgdorferi strain 297 is shown (WT) with the dIR (solid arrows) and the overlapping pIR (dashed arrows). The nucleotides that have been changed are marked in bold. The strain nomenclature is as follows: dIR+ has the nucleotide sequence changed but the complementarity of the inverted repeats maintained, and dIR− has the distal inverted repeat disrupted but the complementarity of the proximal IR intact. (B) PCR of genomic DNA from ospC operator mutants (lane 1, WT with flgBp-aphI cassette; lane 2, dIR+; lane 3, dIR−; and lane 4, no template control) using primers primers kanR 488R (a) and ospC D1572R+AgeI (b) to determine the orientation of the flgBp-aphI antibiotic resistance cassette.