TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| S17-1λpir | Recipient for suicide plasmids | 48 |
| BL21(DE3) | Host for protein expression experiments | 51 |
| E. cloni | Recipient for cloning experiments | Lucigen |
| TOP10 | Recipient for cloning experiments | Invitrogen |
| Y. pseudotuberculosis | ||
| SAL1 | YPIII(pIB102), wild-type parental strain | 7 |
| SAL2 | yscP mutant | This study |
| SAL3 | yscP yscU double mutant | This study |
| SAL4 | yscI mutant | This study |
| SAL5 | yscS mutant | 5 |
| SAL6 | yscI yscP double mutant | This study |
| Plasmids | ||
| pSE380 | High-copy-no. expression vector | Invitrogen |
| pSW71 | Vector for YscI expression from pSE380 | This study |
| pSW71-P77A | P77A mutation in pSW71 | This study |
| pSW71-L80A | L80A mutation in pSW71 | This study |
| pSW71-M83A | M83A mutation in pSW71 | This study |
| pSW71-Q84A | Q84A mutation in pSW71 | This study |
| pSW71-W85A | W85A mutation in pSW71 | This study |
| pSW71-L87A | L87A mutation in pSW71 | This study |
| pSW71-R89D | R89D mutation in pSW71 | This study |
| pSW71-I90A | I90A mutation in pSW71 | This study |
| pSW71-T91A | T91A mutation in pSW71 | This study |
| pSW71-Q93A | Q93A mutation in pSW71 | This study |
| pSW71-E94K | E94K mutation in pSW71 | This study |
| pSW71-E95K | E95K mutation in pSW71 | This study |
| pSW71-L96A | L96A mutation in pSW71 | This study |
| pSW71-I97A | I97A mutation in pSW71 | This study |
| pSW71-A98V | A98V mutation in pSW71 | This study |
| pSW71-K99E | K99E mutation in pSW71 | This study |
| pSW71-T100A | T100A mutation in pSW71 | This study |
| pSW71-G102A | G102A mutation in pSW71 | This study |
| pSW71-Q106A | Q106A mutation in pSW71 | This study |
| pSW71-N107A | N107A mutation in pSW71 | This study |
| pSW71-E109K | E109K mutation in pSW71 | This study |
| pSW71-T110A | T110A mutation in pSW71 | This study |
| pSW71-L111A | L111A mutation in pSW71 | This study |
| pSW71-K113E | K113E mutation in pSW71 | This study |
| pBAD18 | Arabinose-inducible PBAD promoter | 18 |
| pBAD33 | PBAD promoter; p15a ori | 18 |
| pSL327 | yscP gene in pBAD18 | This study |
| pJJ8 | yscU gene in pBAD18 | This study |
| pJJ10 | yscP gene in pBAD33 | This study |
| pSW30 | yscU(Y287G) gene in pBAD18 | This study |
| pSW31 | yscU(Y317D) gene in pBAD18 | This study |
| pGEX-GP-1 | GST fusion vector; parent vector | GE Healthcare |
| pSL303 | GST-YscF overexpression vector | This study |
| pSL310 | GST-YscI overexpression vector | This study |
| pDM4 | Suicide vector | 35 |
| pSL217 | yscP deletion in pDM4 | This study |
| pJJ1 | yscI deletion in pDM4 | This study |
| pJJ2 | yscU deletion in pDM4 | This study |