TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus | ||
| RN4220 | NCTC8325-4 derivative, antibiotic susceptible, restriction deficient; sau1 hsdR (rK− mK+) | 49 |
| RN4220/pVPF5 | RN4220 containing plasmid pVPF5 from S. xylosus VF5 | 43 |
| E. coli | ||
| DH5α | K-12 strain; recA1 endA1 hsdR17 (rK− mK+) | Life Technologies |
| AG100 | K-12 strain | 50 |
| AG100A | AG100 ΔacrAB::aphA1 (Kmr) mutant; increased susceptibility to antibiotics | 44 |
| B. subtilis | ||
| DSM4514 | Derivative of strain 168 containing plasmid pUB110 | DSMZ |
| Plasmids | ||
| pUB110 | S. aureus plasmid, replicates in B. subtilis; aadD (Kmr Neor) | 51 |
| pVPF5 | S. xylosus pT181 family plasmid; tet(K) | 43 |
| pBUS1 | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon, ColE1, MCS pBluescript II SK (Stratagene), (rrnB T1)5 tet(L) | 24 |
| pBUS1-Pcap | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon, ColE1, promoter of S. aureus type 1 capsule gene 1A (Pcap) upstream of a new MCS with joinable rgs-his6 codons (Pcap-MCS-rgs-his6), (rrnB T1)5 tet(L) | This study |
| pBUS1-HC | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon, sso oriL ColE1, MCS pBluescript II SK (Stratagene), (rrnB T1)5 tet(L) | This study |
| pBUS1-Pcap-HC | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon, sso oriL ColE1 Pcap-MCS-rgs-his6 (rrnB T1)5 tet(L) | This study |
| pTSSC | S. aureus-E. coli shuttle vector; pT181 replicon, ColE1, MCS pBluescript II SK (Stratagene), (rrnB T1)5 tet(L) | This study |
| pTSSC-Pcap | S. aureus-E. coli shuttle vector; pT181 replicon, ColE1 Pcap-MCS-rgs-his6 (rrnB T1)5 tet(L) | This study |
| pTSSCm | pTSSC with two silent mutations in the repC sequence to remove HindIII and XbaI sites | This study |
| pTSSCm-Pcap | pTSSC-Pcap with two silent mutations in the repC sequence to remove HindIII and XbaI sites | This study |
| pBJW13 | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon; ColE1 Pcap erm(44) (MLSBr) (rrnB T1)5 tet(L) | 26 |
| pBJW13-HC | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon, sso oriL ColE1 Pcap erm(44) (rrnB T1)5 tet(L) | This study |
| pTJW13 | S. aureus-E. coli shuttle vector; pT181 replicon, ColE1 Pcap erm(44) (rrnB T1)5 tet(L) | This study |
| pBJW13-RGS-His | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon, ColE1 Pcap erm(44)-rgs-his6 (rrnB T1)5 tet(L) | This study |
| pBJW13-HC-RGS-His | S. aureus-E. coli shuttle vector; pAMα1 minimum replicon, sso oriL ColE1 Pcap erm(44)-rgs-his6 (rrnB T1)5 tet(L) | This study |
| pTJW13-RGS-His | S. aureus-E. coli shuttle vector; pT181 replicon, ColE1 Pcap erm(44)-rgs-his6 (rrnB T1)5 tet(L) | This study |