TABLE 1.
Bacterial strains and plasmid constructs used in this study
| Strain or plasmid | Commentsa | Reference(s) or source |
|---|---|---|
| B. burgdorferi strains | ||
| MSK5 | B. burgdorferi strain B31; contains all known plasmids | 47 |
| ML23/pBSVφ(flaBp-gfp) | B. burgdorferi strain B31 lacking only lp25; expresses the cycle 3 gfp allele | 14, 47 |
| ML23/pEW159 | B. burgdorferi strain B31 lacking only lp25; expresses the cycle 3 gfp allele and contains bbe22 | This study |
| JS315/pEW159 | ML23 derivative carrying bbk32::Strr; expresses the cycle 3 gfp allele and contains bbe22 | This study |
| ML23/pJW201 | B. burgdorferi strain B31 lacking only lp25; expresses the gfp mut1 allele and contains bbe22 | This study |
| JS315/pJW201 | ML23 derivative carrying bbk32::Strr; expresses the gfp mut1 allele and contains bbe22 | This study |
| JS315/pJW202 | ML23 derivative carrying bbk32::Strr; expresses the gfp mut1 allele and contains bbe22 and bbk32 | This study |
| E. coli strains | ||
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rK− mK+) supE44 thi-1 gyrA96 relA1 λ− | Invitrogen |
| Plasmids | ||
| pBSVφ(flaBp-gfp) | Borrelial shuttle vector containing constitutively expressed cycle 3 gfp allele; Kanr | 14 |
| pBBE22 | Borrelial shuttle vector carrying bbe22; Kanr | 67 |
| pCR8/GW/TOPO | Gateway PCR cloning/entry vector; Spcr | Invitrogen |
| pJW101 | pCR8/GW/TOPO containing constitutively expressed cycle 3 gfp allele from pBSVΦ(flaBp-gfp); Spcr | This study |
| pBBE22gate | pBBE22 modified into a Gateway destination vector containing attR sites; Camr Kanr | 87 |
| pEW159 | Constitutively expressed cycle 3 gfp allele recombined via Gateway system into pBBE22gate; Kanr | This study |
| pTM61 | Modified borrelial shuttle vector pTM49 that contains the gfp mut1 allele; Genr | 59 |
| pJW201 | bbe22 region cloned into pTM61; Genr | This study |
| pCR2.1/BbrecA | B. burgdorferi recA cloned into pCR2.1-TOPO; Kanr Ampr | This study; D. Shaw |
a
Camr, chloramphenicol resistance; Genr, gentamicin resistance; Strr, streptomycin resistance; Ampr, ampicillin resistance; Kanr, kanamycin resistance; Spcr, spectinomycin resistance.