TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Reference |
|---|---|---|
| Strains | ||
| Kingella kingae | ||
| 269-492 | K. kingae isolate from St. Louis Children's Hospital | 13 |
| KK01 | Stable nonspreading/noncorroding derivative of 269-492 | 13 |
| ctrA | KK01 with the aphA3 (Kanr) cassette inserted into ctrA, resulting in inactivation of ctrABCD | 3 |
| lipA | KK01 deletion of lipA, containing the aphA3 cassette | This study |
| lipB | KK01 deletion of lipB, containing the aphA3 cassette | This study |
| csaA | KK01 deletion of csaA, containing the aphA3 cassette | This study |
| lipA(lipA) | Inducible complement of lipA in the lipA background | This study |
| lipB(lipB) | Inducible complement of lipB in the lipB background | This study |
| csaA(csaA) | Inducible complement of csaA in the csaA background | This study |
| lipA pilA1 | lipA with the cat (Cmr) cassette disrupting pilA1, resulting in inactivation of pilA1 | This study |
| lipB pilA1 | lipB with a cat cassette disrupting pilA1, resulting in inactivation of pilA1 | This study |
| csaA pilA1 | csaA with a cat cassette disrupting pilA1, resulting in inactivation of pilA1 | This study |
| lipA(lipA) pilA1 | lipA(lipA) with a cat cassette disrupting pilA1, resulting in inactivation of pilA1 | This study |
| lipB(lipB) pilA1 | lipB(lipB) with a cat cassette disrupting pilA1, resulting in inactivation of pilA1 | This study |
| csaA(csaA) pilA1 | csaA(csaA) with a cat cassette disrupting pilA1, resulting in inactivation of pilA1 | This study |
| CsaAD99A | Amino acid change in the first D residue of the DXD motif | This study |
| CsaAD101A | Amino acid change in the second D residue of the DXD motif | This study |
| CsaAA99D* | Reversion of CsaAD99A to wild type | This study |
| CsaAA101D* | Reversion of CsaAD101A to wild type | This study |
| CsaAH518A | Amino acid change of the H residue of the HP motif | This study |
| CsaAP519A | Amino acid change of the P residue of the HP motif | This study |
| CsaAA518H* | Reversion of CsaAH518A to wild type | This study |
| CsaAA519P* | Reversion of CsaAP519A to wild type | This study |
| E. coli | ||
| DH5α | F− ϕ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ− thi-1gyrA96 relA1 | 40 |
| BL21(DE3) | F− ompT hsdSB(rB− mB−) gal dcm (DE3) | 41 |
| Plasmids | ||
| pUC19ctrA | ctrA disruption construct marked with the aphA3 cassette | 3 |
| pUC19lipA | lipA disruption construct marked with aphA3 | This study |
| pTrcCErmlipAcomp | Complementation vector containing full-length lipA | This study |
| pUC19lipB | lipB disruption construct marked with aphA3 | This study |
| pTrcCErmlipBcomp | Complementation vector containing full-length lipB | This study |
| pUC19csaA | csaA disruption construct marked with aphA3 | This study |
| pTrcCErmcsaAcomp | Complementation vector containing full-length csaA | This study |
| pTrcCErm | Complementation vector (pUC19) containing the trc promoter that targets recombination to complementation locus with homologous flanking regions; contains the ermC (Ermr) cassette | This study |
| pCsaA5′ | pUC19 containing the coding sequence for the CsaA N terminus and upstream 5′ sequence | This study |
| pCsaAD99A | pCsaA5′ containing a D99A point mutation | This study |
| pCsaAD101A | pCsaA5′ containing a D101A point mutation | This study |
| pCsaAA99D* | pCsaA5′ containing an A99D reversion | This study |
| pCsaAA101D* | pCsaA5′ containing an A101D reversion | This study |
| pCsaA3′ | pUC19 containing the coding sequence for the CsaA C terminus and downstream 3′ sequence | This study |
| pCsaAH518A | pCsaA5′ containing an H518A point mutation | This study |
| pCsaAP519A | pCsaA5′ containing a P519A point mutation | This study |
| pCsaAA518H* | pCsaA5′ containing an A518H reversion | This study |
| pCsaAA519P* | pCsaA5′ containing an A519P reversion | This study |
| pNtermCsaA | pET-15b containing csaA inserted into XhoI/BamHI sites | This study |
| pIND4 | Source of the ermC erythromycin resistance gene | 42 |
| pACYC184 | Source of the cat chloramphenicol resistance gene | 43 |
| pFalcon2 | Source of the aphA3 kanamycin resistance gene | 44 |
| pUC19pilA1 | pilA1 disruption construct marked with cat | This study |