TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| One Shot, TOP10 | Competent E. coli for plasmid transformation | Invitrogen |
| CYL316 | RN4220, pYL112Δ19, L54 int gene, r− | 27 |
| RN6390 | Derivate of NCTC 8325-4 | 36 |
| RN6112 | RN6390 agr::Tn551 | Richard Novick |
| 8325 (ΔsigB) | ΔrsbVW-sigB::erm | 24 |
| RN4220 | Restriction-deficient S. aureus strain | 23 |
| RN4220-29 | RN4220 Δsae::kan | Unpublished data |
| Newman | Wild type | 7 |
| Newman-29 | Newman, Δsae::kan | This study |
| Newman-agr | Newman, agr::Tn551 | This study |
| ISP479R | ISP479R, derivative of strain 8325-4, in which the mutation in rsbU was repaired to gain full sigma B activity | 40 |
| ISP479R-29 | Δsae::kan | This study |
| ISP479R-agr | agr::Tn551 | This study |
| ISP479R-sigB | IS479R, ΔrsbVW-sigB::erm | This study |
| UAMS-1 | Osteomyelitis isolate | 9 |
| UAMS-1-agr | agr::Tn551 | This study |
| UAMS-1-sigB | UAMS-1, ΔrsbVW-sigB::erm | This study |
| Plasmids | ||
| pDG783 | Vector containing kanamycin resistance cassette | 18 |
| pKO010 | Cloning vector with spoVG-lacZ | 34 |
| pCL84 | Single-copy integration vector, att-site for chromosomal integration in geh | 27 |
| pCG6 | pCL84 with integrated spoVG-lacZ gene cassette from pKO010 | This study |
| pCG7 | pCG6 with sae P1 (nt 1 to 400) | This study |
| pCG8 | pCG6 with sae P2 + short (nt 496 to 1141) | This study |
| pCG38 | pCG6 with sae P2 + middle (nt 496 to 1258) | This study |
| pCG39 | pCG6 with sae P2 + long (nt 496 to 1269) | This study |
| pCG9 | pCG6 with sae P3 + short (nt 1127 to 1529) | This study |
| pCG36 | pCG6 with sae P3 + middle (nt 974 to 1529) | This study |
| pCG15 | pCG6 with sae P3 + long (nt 496 to 1529) | This study |
| pCWSAE28 | pCL84 with saePQRS operon (long, nt 1 to 3515) | This study |
| pCWSAE47 | pCL84 with partial sae operon including P3 (short, nt 1066 to 3515) | This study |
| pCWSAE48 | pCL84 with partial sae operon including P2 and P3 (middle, nt 724 to 3515) | This study |