TABLE 1.
Summary of bacterial strains, bacteriophage, and plasmids used in this study
| Strain, phage, or plasmid | Genotype, phenotype, description | Reference or source |
|---|---|---|
| E. coli DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF)U196 endA1 recA1 hsdR17(rK− mK+) deoR thi-1 supE44 λ− gyrA96 relA1 | BRL |
| S. aureus | ||
| 8325-4 | Wild-type strain 8325 UV cured of phages φ11, φ12, and φ13 | National Culture Type Collection |
| PM466 | agr-null mutant of RN6390 | This study |
| PM614 | PM466 chr::Tn917::rot φ11 transductant | This study |
| PM615 | PM466 chr::Tn917::rot φ11 transductant | This study |
| PM616 | PM466 chr::Tn917::rot φ11 transductant | This study |
| PM702 | RN6390 chr::Tn917::rot φ11 transductant | This study |
| PM720 | PM614 with rot restored by allelic exchange | This study |
| RN4220 | 8325-4, nitrosoguanidine-induced restriction mutant used as primary recipient for plasmids propagated in E. coli | |
| RN6390 | 8325-4 | 32 |
| RN6911 | 8325-4 Δagr | 33 |
| Phage φ11 | S. aureus generalized transducing phage | |
| Plasmids | ||
| pBC SK | Cloning vector | Stratagene |
| pBluescript SK(+) | Cloning vector | |
| pCRII (+) | t-tail cloning vector | Invitrogen |
| pJM33 | pRN6650 with a 3.3-kb Agr-encoding ClaI-HpaI deletion | This study |
| pJM36 | pBluescript SK(+) with a 1.2-kb Tn551 transposase-encoding HindIII fragment | 27 |
| pJM37 | pBluescript KS with a 1.2-kb HindIII insert that encodes the transposase of Tn917 | This study |
| pJM42 | pT7::rnaiii; RNA III gene from RN6390 amplified using primers 5′-CACAGAGATGTGATGGAAAATAG and 5′-CATGACTAAACATAGATTTATGAG | This study |
| pJM48 | pSPT181(ts)::agr-null | This study |
| pJM202 | pSPT181(ts)::rot | This study |
| pJM531 | pCR-Script::hla; alpha-toxin gene from RN6390 amplified using primers 5′-GGAAGCTTAAACATCATTTCTGAAGTTATCGGC and 5′-GGGACTAGTGAAGGATGATGAAAATGAAAACACG | This study |
| pRN6650 | pUC18::agr; contains a 6.1-kb MboI Agr-encoding fragment | 35 |
| pSPT181(ts) | Temperature-sensitive S. aureus-E. coli shuttle vector | 17 |
| pT7Blue | t-tail vector | Novagen |
| pTV1 | E. coli-S. aureus shuttle vector with Tn917 | 41 |