TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Reference or source |
|---|---|---|
| E. faecalis JH2-2 | Plasmid-free derivative of E. faecalis JH-2 | 49 |
| E. coli DH5α | supE44 ΔlacU169(φ80lacZΔM15)hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Bethesda Research Laboratories |
| E. coli RYC1000 | araD139 ΔlacU169 rpsL Δrib7 thiA gyrA recA56 | 7 |
| pAM401 | Bifunctional E. coli-E. faecalis cloning vector; Cmr Tcr | 17 |
| pMB2 | AS-48+ Imm+; 68-kb as-48 cluster (as-48ABCC1DD1EFGH) | 32 |
| pMG36e | Lactoccocal cloning and expression vector, replicative in E. coli; Eryr | 48 |
| pGEM-3Zf(+) | Cloning vector; Ampr | Promega |
| pSL1180 | Cloning vector; Ampr | Pharmacia |
| pAM401-76 | AS-48−, partially Imm+; 1.9-kb SphI-BglII (as-48A promoter (PA) and as-48A structural gene) and 6.6-kb BglII (as-48H) DNA fragments from pMB2 cloned into pAM401; Cmr | 12 |
| pAM401-81 | AS-48+ Imm+; complete 25-kb as-48 region (PA and as-48ABCC1DD1EFGH) from pMB2 cloned into pAM401; Cmr | 12 |
| pAM401-52 | AS-48+ partially Imm+; 7.8-kb SphI-PstI (PA and as-48ABCC1DD1EF) DNA fragment from pMB2 cloned into pAM401; Cmr | 34 |
| pAM401-58 | AS-48− and Imm+ 0.16.7 kb of the as-48 region (carries the 3′ region of as-48A, the intergenic region between as-48A and as-48B, and as-48BCC1DD1EF); derivative of pAM401-52 by deletion of the 1.7-kb SphI-PstI DNA fragment; Cmr Tcr | 34 |
| pAM401-81:: Tn5PA | Tn5 insert in PA (as-48A promoter); Cmr Tcr | This work |
| pAM401-81::Tn5B | Tn5 insert in as-48B gene; Cmr Tcr | This work |
| pAM401-81::Tn5C | Tn5 in as-48C gene; Cmr Tcr | This work |
| pMG36-A | PA-as-48A cloned into pMG36e; Eryr | This work |
| pMG36-BC | as-48BC cloned into pMG36e under control of P32 promoter; Eryr | This work |
| pAM401-76X | pAM401-76 containing XhoI restriction site 22 nt downstream of as-48A gene; Cmr | This work |
| pBgD12S | 1.9-kb SphI-BglII D fragment from pMB2 cloned into pSL1180; Ampr | This work |
| pAM401-81X | pAM401-81 containing XhoI restriction site 22 nt downstream of the as-48A gene; Cmr | This work |
a
Cmr, chloramphenicol resistant; Tcr, tetracycline resistant; Ampr, ampicillin resistant; Eryr, erythromycin resistant; AS-48+, AS-48 production; AS-48−, no AS-48 activity detected; AS-48r, AS-8 resistant; AS-48s, AS-48 sensitive.