TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or annotation | Source, reference, or derivation |
|---|---|---|
| Strains | ||
| E. coli | ||
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL endA1 nupG | Invitrogen |
| χ6212 | asd-negative DH5α derivative | 31 |
| Y. pestis | ||
| KIM6+ | Pgm+ pMT1 pPCP1, cured of pCD1 | 16 |
| KIM5+ | Pgm+ pMT1 pPCP1 pCD1Ap | 16 |
| KIM5 | Pgm− pMT1 pPCP1 pCD1Ap | 5 |
| χ10010 | Δcrp-18 Y. pestis KIM6+ | This study |
| χ10017 | ΔPcrp21::TT araC PBADcrp Y. pestis KIM6+ | This study |
| Plasmids | ||
| pUC18 | Apr | Invitrogen |
| pKD46 | repA101(Ts) ori λ Red recombinase expression plasmid | 12 |
| pYA3493 | Asd+ pBR ori β-lactamase signal sequence-based periplasmic secretion plasmid | 21 |
| pYA3700 | TT araC PBAD cassette plasmid, Apr | 9 |
| pYA4373 | cat-sacB cassette in the PstI and SacI sites of pUC18 | 48 |
| pYA4443 | Six-His tag in the C-terminal end of lcrV gene cloned into EcoRI and HindIII sites of pYA3493 | pYA3493 |
| pYA4579 | y3957′-′y3955 fragment ligated by overlapping PCR cloned into EcoRI and HindIII sites of pUC18 | pUC18 |
| pYA4581 | SD-crp and y3957′ fragments cloned into XhoI/EcoRI sites and PstI/HindIII sites of pYA3700 | pYA3700 |
| pYA4588 | cat-sacB cassette from pYA4373 cloned into PstI site of pYA4581 | pYA4581 |