Table 1.
Bacterial strains and plasmids used in this studya
| Strain or plasmid | Relevant feature(s) | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH5α | Cloning host | Lab stock |
| E. faecalis | ||
| OG1Sp | Spr | 21 |
| 100-5 | Spr; carries chromosomal copy of prgX | 14 |
| DM105 | Fusr; rpoC::His allele | This study |
| B. subtilis | rpoC::His | 26 |
| Plasmids | ||
| pGEM-T Easy | Crbr; TA cloning vector | Promega |
| pCF10 | Tcr; cCF10 conjugative plasmid | 12 |
| pCI3340 | Camr; vector used for all plasmids below | 17 |
| pBK1 | Camr | |
| pBK2 | Camr | |
| pDM1 | pCI3340 with prgX terminator | This study |
| pDM1.1 | pDM1 carrying LT −163 to +114 | This study |
| pDM1.2 | pDM1 carrying LT with B1m1 mutation | This study |
| pDM1.3 | pDM1 carrying LT with B1m2 mutation | This study |
| pDM1.4 | pDM1 carrying LT with B2m1 mutation | This study |
| pDM1.5 | pDM1 carrying LT with B2m2 mutation | This study |
| pDM1.6 | pDM1 carrying LT with B1m1 and B2m1 mutations | This study |
| pDM1.7 | pDM1 carrying ST −68 to +114 | This study |
| pDM1.8 | pDM1 carrying ST with B2m1 mutation | This study |
a
The pDM1.1 to -1.8 plasmids contain cloned pCF10-derived DNA fragments that were also used as templates for in vitro transcription. The sequences of the PrgX binding sites, as well as those of the B1 and B2 mutations analyzed in these studies, are shown in Fig. 1C.