TABLE 1.

Strain and plasmids used in this study

Strain or plasmidDescriptionReference
E. faecalis strains
    OG1SSpStreptomycin- and spectinomycin-resistant wild-type strain6
    TX5128OG1RF derivative, gelEsprE27
Plasmids
    pCF10Pheromone-inducible conjugative plasmid5
    pMSP3535Cloning shuttle vector with nisin-inducible promoter2
    pMSP3609Double RGD → RAD mutations in prgB inserted into the pMSP3535 vector32
    pCF10-1Asc10 mutant derivative; 31-amino-acid insertion in central aggregation domain at nucleotide 163812
    pCF10-2Asc10 mutant derivative; double RGD mutant with RGD → RAD single-amino-acid substitutionsThis study
    pCF10-3Asc10 mutant derivative; N-terminal single RGD mutant (RGDS → RADS)This study
    pCF10-4Asc10 mutant derivative; N-terminal aggregation domain deletion (nucleotides 468 to 1074)This study
    pCF10-5Asc10 mutant derivative; double aggregation domain mutant; 31-amino-acid insertion in central aggregation domain and deletion in N-terminal aggregation domainThis study
    pCF10-6Asc10 mutant derivative; C-terminal domain deletion (nucleotides 2064 to 3414)This study
    pCF10-7Asc10 mutant derivative; C-terminal single RGD mutant (RGDV → RADV)This study
    pCF10-8prgB deletion mutant retaining first three and last three codons, Asc10This study
    pCWΔ468-1074pMSP3535 with in-frame deletion of nucleotides 468 to 1074 in prgB34
    pCWΔ468-1074/Ω1638pMSP3535 carrying double aggregation domain mutant34
    pCWΔ2064-3414pMSP3535 with in-frame deletion of nucleotides 2064 to 3414 in prgB34
    CK104OG1RFΔupp12
    pVE6007Supplies RepA (temperature sensitive), Cmr12
    pCJK2P-upp expression cassette cloned into pORI28012
    pCJK47Conjugative donor plasmid, carries oriTpCF10 and P-pheS*; pORI280 derivative; Emr11
    pCJK47ΔprgBprgB deletion construct with first 9 and last 9 nucleotides, cloned into pCJK47This study
    pCJK2-2prgB double RGD mutant cloned into pCJK2This study
    pCJK2-4prgB in-frame deletion of nucleotides 468 to 1074 cloned into pCJK2This study
    pCJK2-5prgB double aggregation domain Δ468-1074/Ω1638 cloned into pCJK2This study
    pCJK2-6prgB in-frame deletion of nucleotides 2064 to 3414 cloned into pCJK2This study
    pCJK2-7prgB C-terminal single RGD mutant cloned into pCJK2This study