TABLE 1.
Bacterial strains and plasmidsa
| Strain or plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| E. coli | ||
| DH5α | Cloning host | 1 |
| DH5α (λpir) | Strain for propagating plasmids with R6K origins, derived from DH5α | S. C. Straley |
| Y. pestis | ||
| KIM6+ | Pgm+ (Hms+ Ybt+) Lcr− | 24 |
| KIM6-2046.1 | Hms+ Ybt− (irp2::kan2046.1) Lcr− Kmr | 47 |
| KIM6-2046.3 | Hms+ Ybt− (Δirp2-2046.3) Lcr− | 2 |
| KIM6-2070.1 | Hms+ Ybt− (ybtS::kan2070.1) Lcr− Kmr | 29 |
| KIM6-2071 | Hms+ Ybt− (ΔybtU2071) Lcr− | This study |
| KIM6-2072 | Hms+ Ybt− (ΔybtT2072) Lcr− | This study |
| Plasmids | ||
| pACYC184 | 4.2-kb cloning vector; Cmr Tcr | 16 |
| pBluescript II KS+ | 2.9-kb cloning vector; Apr | Stratagene |
| pEUPP1 | 15.4-kb low-copy-number psn::lacZ reporter plasmid; Spcr; iron-, Fur-, and YbtA-regulated expression of β-galactosidase | 22 |
| pEUYbtP | 15.4-kb low-copy-number ybtP::lacZ reporter plasmid; Spcr; iron-, Fur-, and YbtA-regulated expression of β-galactosidase | 23 |
| pKNG101 | 6.8-kb suicide vector; sacB+; R6K origin; Smr | 39 |
| pPROEX-1 | 4.7-kb protein expression vector; Apr | Gibco/BRL |
| pPSN3 | 9.9-kb SalI fragment from pSDR498.4 ligated into pBGL2; Apr | 25 |
| pPSN345 | 3.45-kb SphI/EcoRI fragment from pPSN3 ligated into pUC18; ybtU+ ybtT+; Apr | This study |
| pSDR498.1 | ∼4.6-kb BamHI/EcoRI fragment from pSDR498 ligated into pHC79; ybtS+; Apr | 29, 35 |
| pSUC1 | 4.7-kb suicide vector; sacB+; R6K origin; Apr | 23 |
| pUC18 | 2.7-kb cloning vector; Apr | 68 |
| pYbtS-H6 | 1,305-bp PCR fragment from pSDR498.1 ligated into pPROEX-1; IPTG-regulated expression of YbtS-H6; Apr | This study |
| pYbtT-H6 | 800-bp PCR fragment from pPSN345 ligated into pPROEX-1; IPTG-regulated expression of YbtT-H6; Apr | This study |
| pYbtU-H6 | 1,100-bp PCR fragment from pPSN345 ligated into pPROEX-1; IPTG-regulated expression of YbtU-H6; Apr | This study |
| pYbtTU1 | ∼3.9-kb SphI fragment containing ybtT and ybtU from pPSN3 ligated into pACYC184; Cmr | This study |
| pYbtT1 | pYbtTU1 with 438-bp PvuI fragment deleted; Cmr | This study |
| pYbtT1.1 | 3.5-kb SphI fragment from pYbtT1 ligated into pSUC1; ΔybtT2072 (deletion of 808 bp), sacB+; R6K origin; Apr | This study |
| pYbtU1 | 526-bp EagI/EcoRV fragment from pYbtTU1 ligated into pBluescript II KS+; Apr | This study |
| pYbtU2 | 467-bp PvuII fragment from pYbtTU1 ligated into the EcoRV site of pYbtU1; Cmr | This study |
| pYbtU2.1 | ∼1.0-kb SacI/SalI fragment from pYbtU2 ligated into pSUC1; ΔybtU2071 (deletion of 225 bp), sacB+; R6K origin; Apr | This study |
| pYbtU2.2 | ∼1.0-kb SalI/SmaI fragment from pYbtU2.1 ligated into pKNG101; ΔybtU (deletion of 225 bp); sacB+; R6K origin; Smr | This study |
a
Y. pestis strains with a plus sign possess an intact 102-kb pgm locus containing the genes for hemin storage (hms) and the Ybt system. All other Y. pestis strains contain a mutation within the pgm locus due to either a deletion or insertion of an antibiotic resistance cassette. Strains synthesizing the siderophore yersiniabactin are designated Ybt+, while those affected in yersiniabactin production are Ybt−. Lcr− indicates the absence of the low-calcium response virulence plasmid pCD1. Apr, Kmr, Spcr, Smr, Tcr, and Cmr, resistance to ampicillin, kanamycin, spectinomycin, streptomycin, tetracycline, and chloramphenicol, respectively.