Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics* | Source or reference |
|---|---|---|
| Y. pestis strains† | ||
| KIM5(pCD1Ap)+ | Apr Pgm+ (Hms+ Ybt+) Lcr+ (pCD1Ap, ′yadA : : bla) Pla+; derived from KIM6+ | Gong et al. (2001) |
| KIM5-2008 (pCD1Ap) | Apr Hms− (hmsH2008 : : mini-kan) Lcr+ (pCD1Ap, ′yadA : : bla) Pla+; derived from KIM6-2008 | This study |
| KIM6+ | Pgm+, (Hms+) Lcr− Pla+ | Fetherston et al. (1992) |
| KIM6 | Pgm− (Δpgm; Hms−) Lcr− Pla+; derived from KIM6+ | Fetherston et al. (1992) |
| KIM6-2008 | Hms− (hmsH2008 : : mini-kan) Lcr− Pla+; derived from KIM6+ | Lillard et al. (1997); Pendrak & Perry (1991) |
| KIM6-2115 | Hms− (in-frame ΔhmsH2115) Lcr− Pla+; derived from KIM6+ | Forman et al. (2006) |
| KIM6-2116 | Hms– (non-polar ΔhmsF2116) Lcr− Pla+; derived from KIM6+ | Forman et al. (2006) |
| KIM6-2118 | Hms− (in-frame ΔhmsR2118) Lcr− Pla+; derived from KIM6+ | Forman et al. (2006) |
| KIM6-2119 | Hms− (in-frame ΔhmsS2119 : : cam) Lcr− Pla+; derived from KIM6+ | Forman et al. (2006) |
| KIM6-2051+ | Hms−, Kmr (hmsT2051 : : mini-kan) Lcr− Pla+; derived from KIM6+ | Kirillina et al. (2004) |
| KIM6-2090.2+ | Pgm+ Hms2+ (ΔhmsP2090.4) Lcr− Pla+; derived from KIM6+ | A. Bobrov and O. Kirillina, University of Kentucky |
| E. coli strains | ||
| DH5α | Cloning strain | Ausubel (1987) |
| DH5α (λ pir) | Strain for maintenance of R6K origin suicide vector | S. C. Straley, University of Kentucky |
| Plasmids | ||
| pNEB193 | 2.7 kb, Apr, cloning vector | New England Biolabs |
| pKNGΔhmsR | 9.0 kb, Smr, 2.2 kb SalI–XbaI fragment from pWSKΔhmsR ligated into SalI–XbaI sites of pKNG101 | Forman et al. (2006) |
| pNPM22 | 9.9 kb, Cmr, hmsH+hmsF′; used in construction of HmsH variant proteins | Lillard et al. (1997); Pendrak & Perry (1993) |
| pNPM22 HmsH-aa substitutions | 9.8 kb, Cmr; HmsH with various amino acid substitutions, 17 in the surface-exposed loops and seven in the predicted N-terminal periplasmic domain | This study |
*Apr, Cmr, Kmr and Smr: resistance to ampicillin, chloramphenicol, kanamycin and streptomycin, respectively. The pgm locus, type III secretion system encoded on pCD1 (Lcr), and plasminogen activator (Pla) are all required for full virulence in Y. pestis.
†Y. pestis strains with a plus sign possess an intact 102 kb pgm locus. Y. pestis Δpgm strains lack the hmsHFRS locus and the yersiniabactin iron transport system.