Table 1.

Strains and plasmids

Strain or plasmidRelevant propertiesReference or source
Strains
E. coli
  DH5αE. coli host strain for pBluescript SK(−)Stratagene
  DH5α(pTCV-lac)E. coli DH5α containing plasmid pTCV-lac20
  TX4577E. coli DH5α containing plasmid pTEX457727
E. faecalis
  OG1RFGel+ serine protease positive (Spr+) Rifr Fusr13
  TX5240OG1RF fsrA mutant with pTEX4577 insertion in fsrA; Gel SprKanr22
  TX5241OG1RF fsrB mutant with pTEX4577 insertion in fsrB; GelSpr Kanr22
  TX5242OG1RFfsrC mutant with pTEX4577 insertion in fsrC; Gel Spr Kanr22
  TX5266OG1RF fsrB deletion mutant, deletion from bp 79 to 684 of fsrB; GelSprThis study
Plasmids
 pBluescript SK(−)Cloning vector; AmprStratagene
 pTEX4577Suicide vector inE. faecalis derived from pBluescript SK(−); Kanr27
 pTCV-lacShuttle vector containing promoterless lacZ; KanrEryr20
 pTEX5267pTEX4577 containingfsrB flanking regions (917-bp 5′ region: bp −839 to +78, amplified using BDF1 and BDR1 primers; 1,065-bp 3′ region: 45 bp before stop codon to 1,020 bp after stop codon, amplified using DBF2 and DBR2 primers), used for construction of fsrB deletion mutant; KanrThis study
 pTEX5268fsrApromoter cloned upstream of lacZ in pTCV-lac, from bp −406 to −6 (401 bp, amplified using APRF1 and APRR1 primers) relative to fsrA start codon; KanrEryrThis study
 pTEX5269fsrBpromoter cloned upstream of lacZ in pTCV-lac, from bp −110 to −8 (103 bp, amplified using BPRF1 and BPRR1 primers) relative to fsrB start codon; KanrEryrThis study
 pTEX5270gelEpromoter cloned upstream of lacZ in pTCV-lac, from bp −218 to −16 (203 bp, amplified using EPRF1 and EPRR1 primers) relative to gelE start codon; KanrEryrThis study
 pTEX5298fsrBpromoter region (bp −90 to −8 relative to fsrB start codon, amplified using BPRF1 and BMP1 primers) cloned upstream oflacZ in pTCV-lac; KanrEryrThis study
 pTEX5299fsrBpromoter region (bp −72 to −8 relative to fsrB start codon, amplified using BPRF1 and BMP2 primers) cloned upstream oflacZ in pTCV-lac; KanrEryrThis study
 pTEX5300fsrBpromoter region (bp −85 to −8 relative to fsrB start codon, amplified using BPRF1 and BMP4 primers) cloned upstream oflacZ in pTCV-lac; KanrEryrThis study
 pTEX5301fsrBpromoter region (bp −91 to −8 relative to fsrB start codon in which bp −70 to −65 were altered from AAGGAA to TTCCTT, amplified using BPRF1 and BMP5 primers) cloned upstream of lacZ in pTCV-lac; Kanr EryrThis study
 pTEX5302fsrB promoter region (bp −72 to −8 relative to fsrB start codon) with putative gelEpromoter regulatory region (bp −188 to −170 relative togelE start codon), amplified using BPRF1 and BMP6 primers, cloned upstream of lacZ in pTCV-lac; Kanr EryrThis study
 pTEX5303gelE promoter region (bp −188 to −16 relative to gelE start codon, amplified using EPRF1 and EMP1 primers) cloned upstream of lacZ in pTCV-lac; Kanr EryrThis study
 pTEX5304gelE promoter region (bp −170 to −16 relative to gelE start codon, amplified using EPRF1 and EMP2 primers) cloned upstream of lacZ in pTCV-lac; Kanr EryrThis study
 pTEX5305gelE promoter region (bp −188 to −16 relative to gelE start codon in which bp −167 to −161 were altered from AAGGAA to TTCCTT, amplified using EPRF1 and EMP5 primers) cloned upstream of lacZ in pTCV-lac; Kanr EryrThis study
 pTEX5306gelE promoter region (bp −170 to −16 relative to gelE start codon) with putative fsrBpromoter regulatory region (bp −91 to −73 relative to fsrBstart codon), amplified using EPRF1 and BMP6 primers, cloned upstream of lacZ in pTCV-lac; KanrEryrThis study