TABLE 1.
Plasmids and strains used in this study
| Plasmid or strain | Descriptiona | Source |
|---|---|---|
| Plasmids | ||
| pGEM-T Easy | TA cloning vector; Ampr | Promega |
| pJSB430A | pGEM-T Easy-bb0238::PflgB-Kan mutation construct; Kanr | This study |
| pJSB201 | pJD7 with the NdeI site in PflgB-aadA mutated; Specr Strepr | This study |
| pJD7 | Borrelial shuttle vector derived from pKFSS1 | 18 |
| pJSB445 | pJSB201::bb0238 ORF complementation vector; Specr Strepr | This study |
| pProEX-HTb | Expression construct; N-terminal, TEV-cleavable His6 tag; Ampr | Life Technologies |
| pJSB482 | pProEX-HTb::codon-adapted bb0238 ORF; no signal peptide; Ampr | This study |
| pJSB568 | pGEM-T Easy::flaB-actin qPCR standard; Ampr | This study |
| pJSB641 | pGEM-T Easy::flaB probe; Ampr | This study |
| pJSB643 | pGEM-T Easy::bb0238 probe; Ampr | This study |
| piRrp2(Kan) | Borrelial shuttle vector with IPTG-inducible rrp2; Kanr | 17 |
| pJSB499 | piRrp2(Kan) with rrp2 replaced by the ebfC ORF; Kanr | This study |
| pJSB598 | pJSB445 with BB0238Y43A mutation; Specr Strepr | This study |
| pJSB599 | pJSB445 with BB0238G47V mutation; Specr Strepr | This study |
| pJSB603 | pJSB445 with BB0238Y50A mutation; Specr Strepr | This study |
| pJSB604 | pJSB445 with BB0238A66G mutation; Specr Strepr | This study |
| pJSB602 | pJSB445 with BB0238P71A mutation; Specr Strepr | This study |
| Strains | ||
| E. coli | ||
| TOP10F′ | F′ [lacIqTn10(Tetr)] mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 nupG ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 | Life Technologies |
| MON1 | F− endA1 recA1 mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15ΔlacX74 araΔ139 Δ(ara-leu)7697 galU galK nupG rpsL λ− | Monserate |
| C41(DE3) | F− ompT hsdSB (rB− mB−) gal dcm (DE3) | Lucigen |
| B. burgdorferi | ||
| Bb297 | Strain 297; infectious human spinal fluid isolate | 14 |
| BbΔbb0238 | Bb297 with bb0238 interrupted with PflgB-Kan marker; Kanr | This study |
| Bbbb0238C | BbΔbb0238 complemented with pJSB445; Kanr Strepr | This study |
| BbiEbfC | Bb297 transformed with pJSB499; Kanr | This study |
| Bbbb0238Y43A | BbΔbb0238 complemented with pJSB598; Kanr Strepr | This study |
| Bbbb0238G47V | BbΔbb0238 complemented with pJSB599; Kanr Strepr | This study |
| Bbbb0238Y50A | BbΔbb0238 complemented with pJSB603; Kanr Strepr | This study |
| Bbbb0238A66G | BbΔbb0238 complemented with pJSB604; Kanr Strepr | This study |
| Bbbb0238P71A | BbΔbb0238 complemented with pJSB602; Kanr Strepr | This study |
a
Spec, spectinomycin; Strep, streptomycin; Amp, ampicillin; Kan, kanamycin.