TABLE 1.
Strain and plasmids used in this study
| Strain or plasmid | Description | Reference |
|---|---|---|
| E. faecalis strains | ||
| OG1SSp | Streptomycin- and spectinomycin-resistant wild-type strain | 6 |
| TX5128 | OG1RF derivative, gelEsprE | 27 |
| Plasmids | ||
| pCF10 | Pheromone-inducible conjugative plasmid | 5 |
| pMSP3535 | Cloning shuttle vector with nisin-inducible promoter | 2 |
| pMSP3609 | Double RGD → RAD mutations in prgB inserted into the pMSP3535 vector | 32 |
| pCF10-1 | Asc10 mutant derivative; 31-amino-acid insertion in central aggregation domain at nucleotide 1638 | 12 |
| pCF10-2 | Asc10 mutant derivative; double RGD mutant with RGD → RAD single-amino-acid substitutions | This study |
| pCF10-3 | Asc10 mutant derivative; N-terminal single RGD mutant (RGDS → RADS) | This study |
| pCF10-4 | Asc10 mutant derivative; N-terminal aggregation domain deletion (nucleotides 468 to 1074) | This study |
| pCF10-5 | Asc10 mutant derivative; double aggregation domain mutant; 31-amino-acid insertion in central aggregation domain and deletion in N-terminal aggregation domain | This study |
| pCF10-6 | Asc10 mutant derivative; C-terminal domain deletion (nucleotides 2064 to 3414) | This study |
| pCF10-7 | Asc10 mutant derivative; C-terminal single RGD mutant (RGDV → RADV) | This study |
| pCF10-8 | prgB deletion mutant retaining first three and last three codons, Asc10− | This study |
| pCWΔ468-1074 | pMSP3535 with in-frame deletion of nucleotides 468 to 1074 in prgB | 34 |
| pCWΔ468-1074/Ω1638 | pMSP3535 carrying double aggregation domain mutant | 34 |
| pCWΔ2064-3414 | pMSP3535 with in-frame deletion of nucleotides 2064 to 3414 in prgB | 34 |
| CK104 | OG1RFΔupp | 12 |
| pVE6007 | Supplies RepA (temperature sensitive), Cmr | 12 |
| pCJK2 | P-upp expression cassette cloned into pORI280 | 12 |
| pCJK47 | Conjugative donor plasmid, carries oriTpCF10 and P-pheS*; pORI280 derivative; Emr | 11 |
| pCJK47ΔprgB | prgB deletion construct with first 9 and last 9 nucleotides, cloned into pCJK47 | This study |
| pCJK2-2 | prgB double RGD mutant cloned into pCJK2 | This study |
| pCJK2-4 | prgB in-frame deletion of nucleotides 468 to 1074 cloned into pCJK2 | This study |
| pCJK2-5 | prgB double aggregation domain Δ468-1074/Ω1638 cloned into pCJK2 | This study |
| pCJK2-6 | prgB in-frame deletion of nucleotides 2064 to 3414 cloned into pCJK2 | This study |
| pCJK2-7 | prgB C-terminal single RGD mutant cloned into pCJK2 | This study |