Table 3.
Oligonucleotides used in this study.
| Oligonucleotide | Sequence (5′ to 3′) | Description |
|---|---|---|
| BamHI-A33us-recF | CCAAGCTTGGTACCGAGCTCGGATCCACACGCGCTAGCATCCTTTAAATT | Primer pair used to amplify 1526 bp region upstream of bba33 with a 22 bp 5′ flanking region homologous to pCR2.1 upstream of the BamHI site and a 3′ flanking region homologous to the 5′ region of the flgB promoter (PflgB) |
| flgB-A33us-recR | TAGGAAATCTTCCACGCCAATTGTTAAAGCTTTCTGTTTCTACGATATCG | |
| aadA-A33ds-recF | GTGAAAAAGTTTAAAAATCAGTTATTCTTCTATAGGTTTTATTTCTG | Primer pair used to amplify the 1527 bp region downstream from bba33 with a 21 bp 5′ flanking region homologous to the aadA 3′ end and a 32 bp 3′ flanking region homologous to sequences downstream of the XhoI site of pCR2.1 |
| A33ds-XhoI-recR | CGAATTGGGCCCTCTAGATGCATGCTCGAGCGGGTGTTGGAATTGGAAACGAACC | |
| flgB-F | CAATTGGCGTGGAAGATTTCC | Primer pair used to amplify PflgB-StrR cassette from pKFSS1 |
| aadA-R | CTGATTTTTAAACTTTTTCACAAATAGG | |
| P1 | GCCAGAGAGCGATATCGTAGAAAC | Primers P1 through P5 were used to confirm the bba33 deletion in B. burgdorferi as well as complementation construct (see Figs 1 and 2). |
| P2 | AATAATAGGTTCATGAAAAATTTGTATG | |
| P3 | CACAAATAGGGTCAAAATTTTGACC | |
| P4 | TGCCCTATCAGAAATAAAACCTATAGAAG | |
| P5 | GAAGTGCCTGGCAGTAAGTTG | |
| A33-NdeI-5′F | ACGCCATATGTATTTAAATGATTTTTCTGGTATG | Primer pair used to amplify bba33 lacking sequences that encode its leader peptide through the stop codon with NdeI (underlined) and BamHI (bold) sites engineered for cloning into pET15b expression vector |
| A33-BamHI-3′R | ACGCGGATCCTTATTTTTGAATTAGTGCTAAAGCAC | |
| A33up210-BamHI-F | ACGCGGATCCGCCAGAGAGCGATATCGTAGAAAC | Primer pair used to amplify a region from 210 bp upstream and 129 downstream of bba33 with BamHI (bold) and SalI (underlined) sites; cloned into pBBE22luc |
| A33dn129-SalI-R | ACGCGTCGACAAATGGCTAGGGTGGAATACAAAC | |
| b-Actin-F | ACGCAGAGGGAAATCGTGCGTGAC | Primer pair used for enumerating copies of mouse β-actin via qPCR (Pal et al., 2008) |
| b-Actin-R | ACGCGGGAGGAAGAGGATGCGGCAGTG | |
| nTM17FrecA | GTGGATCTATTGTATTAGATGAGGCT | Primer pair used for enumerating copies of B. burgdorferi recA via qPCR (Liveris et al., 2002; Weening et al., 2008) |
| nTM17RrecA | GCCAAAGTTCTGCAACATTAACACCT |