TABLE 2.
Plasmids used in this studya
| Host strain and plasmid | Relevant markers and/or phenotype | Properties | Source or reference |
|---|---|---|---|
| E. coli | |||
| pGEM-7Z(f)+ | Apr | Cloning vector | Promega |
| pGN23 | Apr | 2.3-kb NsiI chromosomal fragment of the S. epidermidis tagF locus in pGEM-7Z(f)+ | This study |
| pGN333 | Apr | 333-bp NsiI chromosomal fragment of S. epidermidis tagF in pGEM-7Z(f)+ | This study |
| pGDH3 | Apr | 1.56-kb ClaI-NsiI internal fragment of the S. epidermidis tagF ORF in pGEM-7Z(f)+ | This study |
| pBluescript | Apr | Cloning vector | Stratagene |
| pBN23 | Apr | 2.3-kb insert from pGN23 in pBluescript | This study |
| pBH17 | Apr | 1.77-kb HindIII fragment from pBCP55 in pBluescript | This study |
| pBCP55 | Apr | 5.5-kb ClaI-PstI chromosomal fragment from S. epidermidis with the 3′ region of tagF and adjacent downstream sequence in pBluescript | This study |
| pBSKT13 | Apr | 2.35-kb HindIII Tcr fragment from pT181 in pBluescript | This study |
| pGEX-KG | Apr | GST fusion-protein expression vector | 16 |
| pGEX-2 | Apr | GST-′TagF fusion protein expression vector | This study |
| B. subtilis and/or E. coli | |||
| pHPS9 | Cmr | Shuttle vector for cloning in E. coli and B. subtilis | 17 |
| pFC10 | Cmr | 3.26-kb S. epidermidis tagF locus in pHPS9 | This study |
| pFCTH3 | Cmr | 3.26-kb tagF locus with a 2-bp mutation in the tagF ORF, in pHPS9 | This study |
| pDG268 | Cmr Apr | Integrational plasmid for complementation analysis in B. subtilis | P. Stragier, Institute de biologie physicochemique, Paris, France |
| pSC5 | Cmr Apr | 3.26-kb S. epidermidis tagF locus in pDG268 | This study |
| S. aureus, pT181 | Cmr Tcr | Carries a 2.35-kb HindIII Tcr fragment | 25 |
| S. aureus and/or S. epidermidis | |||
| pTS2 | Cmr, ts rep | Derived from pTV1ts | This laboratory, MSC from pBluescript |
| pTS2T | Cmr Tcr, ts rep | pTS2 containing the 2.35-kb HindIII Tcr fragment from pBSKT13 | This study |
| pTSTAG | Cmr Tcr, ts rep | 1.56-kb internal tagF fragment from pGDH3 cloned in the MCS of pTS2T | This study |
| pTSH17 | Cmr Tcr, ts rep | 1.77-kb fragment from pBH17, cloned in the MCS of pTS2T | This study |
a
MSC, multiple cloning site; ts rep, temperature-sensitive replication system.