Table 1.

Bacterial strains and plasmids used

Strain or plasmidDescriptionReference or source
Strains
E. faecalis
  OG1X str gel 29
  JH2-2 rif fus 32
  FA2-2 rif fus 14
  DS16Clinical isolate; carries pAD1 and pAD2 14
  39-5Clinical isolate; carries pPD1 and 5 other plasmids 51
  FA373FA2-2 carrying pAM373; Tn918(tet) on chromosome 12
  FA3328OG1X eep mutant (OG1X containing integrated pAM3328)This study
  FA3328RRevertant of FA3328 that had lost erm This study
  FA3329OG1X eep mutant (OG1X containing integrated pAM3329)This study
  FA3329RRevertant of FA3329 that had lost erm This study
E. coli DH5αF φ80lacZΔM15 endA1 recA1 hsdR17(rK mK +) supE44 thi-1 ΔgyrA96lacZYA-argF)U169 Promega Research Labs
Plasmids
 pAD160 kb; encodes hemolysin/bacteriocin (cytolysin); responds to cAD1; originally found in DS16 14
 pAM2011pAD1::Tn917lac with insert 4 bp downstream from t2 41
 pAM2011EMiniplasmid derivative representing largeEcoRI fragment of pAM2011 46
 pPD1Encodes bacteriocin; responds to cPD1 pheromone 51
 pOB1Encodes hemolysin/bacteriocin; responds to cOB1 pheromone 39
 pCF10Carries Tn925 (tet); responds to cCF10 pheromone 19
 pAM373Encodes response to cAM373 pheromone 12
 pAM401 E. coli-E. faecalis shuttle; cat tet 50
 pBluescript SK+II E. coli plasmid vector; amp Stratagene
 pVA891 E. coli plasmid vector; erm cat 34
 pAM3325pAM401 derivative with 3,045-bp cloned from E. faecalis OG1XThis study
 pAM3326pBluescript derivative with 3,045-bp fragment subcloned from pAM3325This study
 pAM3327pAM401 derivative with PCR product representing ORF1 (eep)This study
 pAM3328pVA891 derivative with PCR fragment representing internal segment of eep This study
 pAM3329pVA891 derivative with PCR fragment representing internal segment of eep (fragment in opposite orientation compared to pAM3328)This study