TABLE 1.
Bacterial strains and plasmids
| Y. pestis strains | Characteristics* | Reference |
|---|---|---|
| CO92 | Pgm+ pCD1+ pPCP+ Gly− | [49] |
| CO92L | Pgm+ pCD1− pPCP+ Gly− (cured of virulence plasmid pCD1) |
This study |
| CO92L glpD+ | Pgm+ pCD1− pPCP+ Gly− (defective glpD’ allele replaced with functional glpD gene in the chromosome) |
This study |
| KIM 5 (D27) | Pgm− pCD1+ pPCP+ Gly+ | Brubaker collection |
| KIM 6+ | Pgm+ pCD1− pPCP+ Gly+ | Brubaker collection |
| KIM 6+ glpD’ | Pgm+ pCD1− pPCP+ Gly− (functional glpD gene replaced with defective glpD’ allele in the chromosome) |
This study |
| KIM 6+ ΔglpABC |
Pgm+ pCD1− pPCP+ Gly+ (glpABC operon deletion) |
This study |
| KIM 6+ glpD’ΔglpABC |
Pgm+ pCD1− pPCP+ Gly− (functional glpD gene replaced with defective glpD’ allele in the chromosome; glpABC operon deletion) |
This study |
| 195-P2 | Pgm− pCD1+ pPCP+ Gly− | Hinnebusch collection |
| TS | Pgm+ pCD1− pPCP+ Gly− | Brubaker collection |
| Salazar | Pgm− pCD1− pPCP+ Gly− | Brubaker collection |
| EV76H | Pgm+ pCD1− pPCP+ Gly− (derivative of the live plague vaccine strain EV76) |
Brubaker collection |
| M23 | Pgm− pCD1+ pPCP+ Gly− | Brubaker collection |
| Kimberley | Pgm+ pCD1− pPCP+ Gly− | Perry collection |
| PEXU-2 | Pgm+ pCD1+ pPCP+ Gly− | CDC collection |
| E. coli strains | Characteristics | Reference |
| DH5α | F-φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rk−, mk+) gal− phoA supE44 λ− thi-1 gyrA96 relA1 |
Invitrogen (Carlsbad, CA) |
| BW25141 | F−, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), Δ(phoB- phoR)580, λ−, galU95, ΔuidA3::pir+, recA1, endA9(del- ins)::FRT, rph-1, Δ(rhaD-rhaB)568, hsdR514 |
[19] |
| Plasmids | Characteristics | Reference |
| pBluescript SK+ | Cloning vector, ApR | Stratagene (La Jolla, CA) |
| pBBR1Tp | Cloning vector, TpR | ATCC (Manassas, VA) |
| pKD46 | Mutagenesis helper plasmid, source of Lambda Red Recombinase, ApR |
[19] |
| pKD4 | Mutagenesis helper plasmid, source of kan cassette, KmRApR |
[19] |
| pCVD442 | Source of sacB gene, ApR | [50] |
| pKD4_Km-sacB | Cloned sacB from pCVD442 in NgoMIV site of pKD4, source of kan-sacB cassette, KmRApR,SucS |
This study |
| pBGLPD5 | Cloned glpD from KIM 6+ in Hind III and BamHI sites of pBluescript SK+ |
This study |
| pTpGLPFKX6 | Cloned glpFKX from KIM 6+ in XbaI and XhoI sites of pBBR1Tp |
This study |
| pCD1Ap | Virulence plasmid pCD1 of Y. pestis KIM labeled with ApR marker |
Provided by R. Perry |
*
All strains contained a large plasmid pMT, encoding capsular antigen F1 and murine toxin Ymt. Pgm: pigmentation locus, pCD1: virulence plasmid encoding T3SS, pPCP: small plasmid encoding pesticin and plasminogen activator Pla, Gly: aerobic glycerol fermentation. glpD: functional glpD gene of KIM. glpD’: defective glpD gene of CO92. KmR, ApR, TpR, and SucS are markers of resistance to kanamycin, ampicillin, trimethoprim, and sensitivity to sucrose, respectively. kan and sacB are genes for kanamycin resistance and sucrose sensitivity, respectively.