Generation of a
mutant. A. A bamD/bb0324 mutant was generated through homologous recombination by replacing the bamD/bb0324 gene with a streptomycin resistance cassette. Red arrows indicate primers used to confirm insertion of the streptomycin cassette in the borrelial genome. B. PCR amplification resulted in a 1.4 kb amplicon in the wildtype strain while the BamD/BB0324 mutant containing the streptomycin cassette produced a 2.4 kb amplicon, as expected. A PCR reaction with no DNA template (Neg) was included as a negative control. DNA sizes, in kilobase pairs, are indicated at left. C. Whole-cell lysate from the BamD/BB0324 mutant (BamD::strepR) and parental (WT) strains were subjected to immunoblot analysis using BamD/BB0324 specific antibodies (top panel). FlaB antisera was used to ensure equal loading of lysates examined (bottom panel).