E. coli Shuttle Vector Generation

E. coli host strain NEB 5-alpha F’ l^q (New England Biolabs) was exclusively used to generate, store, and amplify the E. coli/B. burgdorferi shuttle vectors listed in Table 1. The resulting strains were grown on LB agar plates or in Super Broth (35 g/L bacto-tryptone, 20 g/L yeast extract, 5 g/L NaCl, and 6 mM NaOH) liquid medium with shaking at 30°C [53 (link)]. Transformation was achieved by heat shock followed by recovery in SOC medium (New England Biolabs) for 1h at 30°C with shaking. Antibiotic selection was achieved using spectinomycin at 50 μg/mL or rifampin at 25 μg/mL in liquid culture or 50 μg/mL in plates.