Table 1.
Plasmids and bacterial strains used in this work
| Plasmid or strain | Relevant feature(s)a | Reference or source |
|---|---|---|
| Plasmids | ||
| pCC1 | 8.1 kb; Cmrori2 oriV | Epicentre |
| pCR2.1-TOPO | 3.9 kb; Kmr | Invitrogen |
| pELS100 | 7.0 kb; shuttle vector E. coli-Lactobacillus; Apr Emr | LMGT |
| pIL252 | 4.6 kb; a LAB vector; Emr | 36 |
| pNO6 | 12.7 kb; shuttle vector E. coli/LAB formed by ligation of pCC1 and pIL252; Cmr Emr | This work |
| pCN37 | 6.3 kb; shuttle vector E. coli-S. aureus; Apr Emr | 8 |
| pT181mcs | 4.8 kb; S. aureus plasmid; Tcr | 2 |
| pNO9 | 12.9 kb; shuttle vector E. coli/S. aureus formed by ligation of pCC1 and pT181mcs; Cmr Tcr | |
| pRJ9 | 10.4 kb; codes for aureocin A53 | 20 |
| pRJ81 | 13.9 kb; Cmr; BamHI/EcoRI-A fragment of pRJ9 cloned into pCC1 | This work |
| pRJ82 | 13.1 kb; BamHI/EcoRI-A fragment of pRJ9 cloned into pELS100 | This work |
| pRJ83 | 10.6 kb; HindIII-A fragment of pRJ9 cloned into pELS100 | This work |
| pRJ84 | 11.0 kb; orf12-orf13-orf14 of pRJ9 cloned into pCC1 | This work |
| pRJ85 | 15.6 kb; orf12-orf13-orf14 cloned into pNO6 | This work |
| pRJ88 | 13.2 kb; orf11-orf10 cloned into pCC1 and ligated to pT181 | This work |
| pRJ89 | 7.0 kb; orf11-orf10 cloned into pCN37 | This work |
| Strains | ||
| E. coli strains | ||
| EPI300 | Epicentre | |
| DH5α | lacZΔM15 hsdR mutant | 33 |
| L. sakei DSM 20017 | Sensitive to aureocin A53 | LMGT |
| M. luteus ATCC 4698 | Sensitive to aureocin A53 | ATCC |
| S. aureus strains | ||
| RN4220 | hsd mutant; sensitive to aureocin A53 | 30 |
| RN9590 | RN4220 carrying pCN37 | 8 |
| A53 | pRJ9; 10.4 kb; Bac+ Imm+ | 20 |
| MB38 | pRJ14 (pRJ9::Tn551); 15.6 kb; Emr | 1 |
| MB139 | pRJ38 (pRJ9::Tn917-lac); 18.9 kb; Emr | 11 |
| MB140 | pRJ39 (pRJ9::Tn917-lac); 18.9 kb; Emr | 11 |
| MB143 | pRJ40 (pRJ9::Tn917-lac); 18.9 kb; Emr | 11 |
| MB145 | pRJ108 (pRJ9::Tn917-lac); 18.9 kb; Emr; low aureocin A53 production | This work |
| MB388 | RN4220 carrying pT181mcs | This work |
| MB420 | RN4220 carrying pRJ88 | This work |
| MB422 | RN4220 carrying pRJ85 and pRJ88 | This work |
| MB425 | RN4220 carrying pNO6 | This work |
| MB426 | RN4220 carrying pRJ85 | This work |
| MB439 | RN4220 carrying pRJ89 | This work |
| MB502 | Plasmidless derivative of strain MB145 | This work |
a
Imm+, presence of immunity to aureocin A53; hsd mutant, deficient in DNA restriction.