TABLE 1.
Strains, plasmids, and oligonucleotides used in this study
| Bacterium, plasmid, or oligonucleotide | Description or sequence | Reference or source |
|---|---|---|
| B. burgdorferi strains | ||
| B31 MI | Medimmune B31; nonclonal, low-passage infectious strain | 14 |
| A3 | Infectious, transformation-competent clone of B31 MI | 23 |
| 6 | Clonal derivative of A3 (WT) | This study |
| 2E6 | ospA mutant generated with pOKOSPAKO (ospA1) | This study |
| 7A | ospA restored, ospB mutant generated with pOKOSPA-C′ (ospA+B1) | This study |
| E. coli strain | ||
| TOP10F′ | Used for propagating plasmids | Invitrogen |
| Plasmids | ||
| pOK12 | P15A replicon, multiple cloning site lacZ′ vector | 67 |
| pTAkanA | B. burgdorferi flaBp-kan cassette in pCR2.1-TOPO | 10 |
| pTAGmA | B. burgdorferi flaBp-aacC1 cassette in pCR2.1-TOPO | 22 |
| pOKLP54 | pOK12 amplicon fused to LP54 amplicon | This study |
| pOKOSPAKO | ospA inactivation plasmid; pOKLP54 SpeI-EcoRI fragment ligated with NheI-EcoRI fragment of pTAkanA | This study |
| pOKOSPA-C′ | ospA restoration plasmid; pOKLP54 BclI-PvuII fragment ligated with BamHI-PvuII fragment of pTAGmA | This study |
| Oligonucleotides | ||
| POK12-F1+AscI | 5′-TGGCGCGCCTCGCCCTTCCCAACAGTTG | This study |
| POK12-B1+AvrII | 5′-ACCTAGGGCGTATTGGAGCTTTCGCG | This study |
| LP54-F2+AscI | 5′-AGGCGCGCCTTGGTGGCAAGGAAAACCATAC | This study |
| LP54-B3+AvrII | 5′-TCCTAGGCCTTTGGCTTGTTGTCTGCG | This study |
| OSP1A | 5′-TAGGATCCAAGCTTAATTAGAACCAAAC | 54 |
| OSP2 | 5′-CTTCCTACACTAGCTGATGC | 54 |
| KANA.5 | 5′-AGCCATATTCAACGGGAAACG | This study |
| KANA.3 | 5′-CAAGTCAGCGTAATGCTCTGCC | This study |
| flaB470F | 5′-TGGTTTGCTCCAACATGAACTC | This study |
| flaB547R | 5′-TGCAAAAATTAACACACCAGCAT | This study |
| flaB523TR | 5′-FAM-ACTTTCAGGGTCTCAAGCGTCTTGGACTTT-TAMRAa | This study |
a
FAM, 6-carboxyfluorescein; TAMRA, 6-carboxytetramethylrhodamine.