Table 3.
Plasmids and E. coli strains used in this study
| Plasmid or E. coli strain | Characteristicsa | Source or reference |
|---|---|---|
| Plasmids | ||
| pCR2.1 TOPO | Cloning plasmid; Kmr | Invitrogen |
| pJQK | Suicide vector for mutagenesis by gene replacement; Kmr | 31 |
| pDONR221 | Cloning plasmid (Gateway Technology); Kmr | This study |
| pMR10 | Destination vector (Gateway Technology); Kmr Cmr | 19 |
| pMM13 | Donor vector, pDONR221 derivative containing a copy of the B. abortuswbkA gene (BAB1_0553); Kmr | This study |
| wbkA-pMM14 | Complementation vector, pMR10 derivative carrying the att fragment from pMM13, Kmr Cmr | This study |
| pMM62 | pCR2.1 TOPO derivative containing a mutant allele of recA(Δ80-283) (BAB1_1224) | This study |
| pMM63 | Mutator plasmid, pJQK derivative containing the BamHI-XbaI fragment of pMM62 | This study |
| pMM72 | pCR2.1 derivative containing F1R4ΔBAB1_0557 fragment generated by PCR overlap | This study |
| pMM73 | Mutator plasmid, pJQK derivative containing the BamHI-XbaI fragment of pMM72 | This study |
| pMM77 | pCR2.1 derivative containing F1R4ΔBAB1_0546 fragment generated by PCR overlap | This study |
| pMM78 | Mutator plasmid, pJQK derivative containing the BamHI-XbaI fragment of pMM77 | This study |
| pMM99 | pCR2.1 derivative containing a fragment of 853 bp from a wbkA deletion intermediate | This study |
| E. coli strains | ||
| TOP10F | F−mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| OmniMAX 2-t1 | F′ [proAB+lacIqlacZΔM15 Tn10(TetR) Δ(ccdAB)] mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80(lacZ)ΔM15 Δ(lacZYA-argF)U169endA1recA1supE44 thi-1 gyrA96relA1tonA panD | Invitrogen |
| S17-1λpir | Mating strain with plasmid RP4 inserted into the chromosome | 35 |
a
Cmr, chloramphenicol resistance; Strr, streptomycin resistance; Kmr, kanamycin resistance.