TABLE 1.

Strains and plasmids used in this study

Strain or plasmidRelevant characteristics or descriptionaSource or reference
Strains
    E. faecalis
        OG1RFReference strain 7, 14
        CK102OG1RF upp1::ermThis study
        CK104OG1RF Δupp2This study
        CK107CK104 ΔsrtA2This study
    E. coli
        DH5α E. coli cloning host
        EC1000 E. coli cloning host, provides RepA in trans 21
Plasmids
    pFW11Cloning vector, source of promoter for P-upp; Spr 31
    pFW14Cloning vector; Cmr 31
    pFW15Cloning vector; Emr 31
    pMSP3535Nisin-inducible expression vector; Emr 4
    pVE6007Supplies RepA(ts); Cmr 22
    pORI280Requires RepA in trans for replication; Emr 21
    pAM401tsTemperature-sensitive shuttle plasmid; Cmr 46
    pCJK2P-upp expression cassette cloned into pORI280This study
    pCJK8Δupp2 allele in pORI280This study
    pCJK11 upp::erm cassette in pAM401tsThis study
    pCJK14ΔsrtA2 allele in pCJK2This study
    pCJK15 E. faecalis srtA cloned into pMSP3535This study
    pCJK2-1 prgBΩ1638 cloned into pCJK2This study
    pCF10Pheromone-inducible conjugative plasmid 6
    pCF10-1pCF10 carrying prgBΩ1638This study
    pCWΩ1638pMSP3535 carrying prgBΩ1638 43
    pCWΩ3599pMSP3535 carrying prgBΩ3599 (lacks cell wall anchor) 43
    pCJK21Spectinomycin resistance gene cloned into XbaI/XhoI sites of pMSP3535; Emr SprThis study
    pCJK22 E. faecalis upp cloned into pCJK21This study
  • a Spr, spectinomycin resistance; Emr, erythromycin resistance; Cmr, chloramphenicol resistance.