TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Reference(s) |
|---|---|---|
| Strains | ||
| Y. pseudotuberculosis 32777 | Serogroup O1 wild-type strain | 38 |
| Y. pseudotuberculosis 32777ΔyopM | ΔyopM; unmarked | This study |
| Y. pseudotuberculosis 32777ΔyopM + pyopMWT | Expresses full-length YopMWT from native promoter | This study |
| Y. pseudotuberculosis 32777ΔyopM + pyopMΔ6-15 | Expresses YopMΔ6-15 from native promoter | This study |
| Y. pseudotuberculosis 32777ΔyopM + pyopMΔ12-C | Expresses YopMΔ12-C from native promoter | This study |
| Y. pseudotuberculosis 32777ΔyopM + pyopMYPTB | Expresses YopMYPTB from native promoter | This study |
| Y. pestis KIM5 | Δpgm; Ampr | 47 |
| Y. pestis KIM5ΔyopM | ΔyopM; unmarked | This study |
| E. coli S17-1λpir | Tpr SmrhsdR pro recA RP4-2-Tc::Mu-Km::Tn7; lysogenized with λpir | 13, 50 |
| E. coli MACH1 | ΔrecA1398 endA1 tonA P80ΔlacM15ΔlacX74 hsdR | Invitrogen |
| E. coli BL21-AI | F−ompT hsdSBgal dcm araB:T7RNAP-tetA | Invitrogen |
| Plasmids | ||
| pSB890 | Suicide vector; Tetr | 34 |
| pFLP2 | Encodes Flp recombinase; sacB; Tetr | 23 |
| pKD4 | Contains npt cassette flanked by FRT sites; Kanr | 12 |
| pSB890-ΔyopMYP | Upstream and downstream regions of yopM in pSB890; Tetr | This study |
| pSB890-yopMYPTB:npt | yopM containing npt cassette flanked by FRT sites; Kanr; Tetr | This study |
| pENTR221-yopM | Full-length Y. pestis yopM open reading frame; Kanr | JCVIa |
| pDEST15 | Creates N-terminal GST fusions; Ampr | Invitrogen |
| pMMB67EH | Broad-host-range expression vector; low copy; Ampr | 18 |
| pDEST-yopMWT | N-terminal GST fusion to full-length YopM; Ampr | This study |
| pDEST-yopMΔ6-9 | Encodes N-terminal GST fusion to YopM lacking LRR6 to LRR9; Ampr | This study |
| pDEST-yopMΔ8-9 | Encodes N-terminal GST fusion to YopM lacking LRR8 and LRR9; Ampr | This study |
| pDEST-yopMΔ10-11 | Encodes N-terminal GST fusion to YopM lacking LRR10 and LRR11; Ampr | This study |
| pDEST-yopMΔ12-13 | Encodes N-terminal GST fusion to YopM lacking LRR12 and LRR13; Ampr | This study |
| pDEST-yopMΔ14-15 | Encodes N-terminal GST fusion to YopM lacking LRR14 and LRR15; Ampr | This study |
| pDEST-yopMΔ6-15 | Encodes N-terminal GST fusion to YopM lacking LRR6 to LRR15; Ampr | This study |
| pDEST-yopMΔ12-C | Encodes N-terminal GST fusion to YopM truncated after LRR11; Ampr | This study |
| pyopMYP | Full-length Y. pestis yopM gene and promoter in pMMB67EH | This study |
| pyopMΔ6-9 | yopMΔ6-9 gene and promoter in pMMB67EH | This study |
| pyopMΔ8-9 | yopMΔ8-9 gene and promoter in pMMB67EH | This study |
| pyopMΔ10-11 | yopMΔ10-11 gene and promoter in pMMB67EH | This study |
| pyopMΔ12-13 | yopMΔ12-13 gene and promoter in pMMB67EH | This study |
| pyopMΔ14-15 | yopMΔ14-15 gene and promoter in pMMB67EH | This study |
| pyopMΔ6-15 | yopMΔ6-15 gene and promoter in pMMB67EH | This study |
| pyopMΔ12-C | yopMΔ12-C gene and promoter in pMMB67EH | This study |
| pyopMYPTB | Full-length Y. pseudotuberculosis yopM gene and promoter in pMMB67EH | This study |
a
JCVI, J. Craig Venter Institute (Y. pestis gateway clone library, Pathogen Functional Genomics Resource Center, 2009).