TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| Strains | ||
| Y. pseudotuberculosis | ||
| YPIII(pIB102) | Wild-type, parental strain; Kmr | 8 |
| YPIII(pIB75) | pIB102 yscU in-frame deletion; Kmr | This study |
| YPIII | Virulence plasmid-cured strain | 7 |
| E. coli | ||
| S17-1λpir | RP4-2 Tc::Mu-Km::Tn7 (λpir) | 14 |
| TOP10 | Commercial One Shot competent cells | Invitrogen |
| BL21(DE3) | IPTG-inducible T7 RNA polymerase | 41 |
| Plasmids | ||
| pCR2.1 | Commercial cloning vector; Ampr | Invitrogen |
| pMMB66EH | Ptac expression vector; Ampr | 16 |
| pET-22b | T7 overexpression vector; Ampr | Novagen |
| pKK223-3 | Ptac expression vector; Ampr | Amersham Pharmacia Biotech |
| pDM4 | Suicide vector carrying sacB; Cmr | 27 |
| pLS13 | 669-bp PCR fragment of ΔyscU25-329 on pDM4 | This study |
| pLS21 | yscU cloned under tac promoter of pMMB66EH | This study |
| pPE33 | yscU cloned under tac promoter of pKK223-3 | Edqvist et al., unpublished |
| pPE40 | yscU(Δ263-266) cloned under tac promoter of pKK223-3 | This study |
| pPE41 | yscU(N263A) cloned under tac promoter of pKK223-3 | This study |
| pPE42 | yscU(P254A) cloned under tac promoter of pKK223-3 | This study |
| pPE43 | yscU(T265A) cloned under tac promoter of pKK223-3 | This study |
| pPE5 | His6yscU(202-354) under T7 promoter of pET-22b | This study |
| pML11 | yscU(211-354) cloned under tac promoter of pMMB66EH | This study |
| pML12 | yscU(211-354)-Flag cloned under tac promoter of pMMB66EH | This study |
| pML13 | yscU-Flag fusion cloned under tac promoter of pMMB66EH | This study |
| pML14 | yscU(264-354) cloned under tac promoter of pMMB66EH | This study |
| pML16 | yscU(Δ263-266) Flag cloned under tac promoter of pMMB66EH | This study |
| pML17 | yscU(N263A) Flag cloned under tac promoter of pMMB66EH | This study |
| pML18 | yscU(P254A) Flag cloned under tac promoter of pMMB66EH | This study |
| pML19 | yscU(T265A) Flag cloned under tac promoter of pMMB66EH | This study |
a
Kmr, Kanamycin resistance; Ampr, ampicillin resistance; Cmr, chloramphenicol resistance.