FIG. 1.

Schematic of the bosR-containing operon and screening of bosR::kan^R mutant, JH300, in B. burgdorferi. A. The schematic depicts the chromosomal region of B. burgdorferi containing bosR and the genetic disruption of bosR with the insertion of P_flgB-kan^R resulting in strain JH300. The arrows indicate the location of oligonucleotide primers utilized to screen clones for presence of kan^R cassette. B. PCR screen of JH300 was performed using primers specific for bosR that amplified a 586 bp product in ML23 or an approximate 2.5-kb product in JH300, indicating the insertion of P_flgB-kan^R. Lane 1: ML23; Lane 2: JH300. Markers (in bp) are shown on the left. C. BosR is not produced in strain JH300. Cultures were grown to 5 × 10⁷/ml and cell lysates were resolved by SDS-PAGE, immobilized on PVDF membranes, and probed with antisera specific for BosR.