TABLE 1.

Strains and plasmids used in this study

Strain or plasmidCharacteristic(s) or originaReference or source
Strains
    E. coli
        DH5αHost strainStratagene, La Jolla, Calif.
        TX5438DH5α with pTEX5438This study
    E. faecalis
        OG1RF (TX4002)bWell-characterized strain; GelE+ SprE+ Rifr FAr 23
        TX5240OG1RF fsrA disruption; GelE SprE Kanr 15
        TX5241OG1RF fsrB disruption; GelE SprE Kanr 15
        TX5242OG1RF fsrC disruption; GelE SprE Kanr 15
        TX5266OG1RF fsrB deletion; GelE SprE 15
        TX5128OG1RF gelE disruption; GelE SprE Kanr 15
        TX5264OG1RF gelE deletion; GelE S-SprE+ 6
        TX5244TX5240 with pTEX5249; GelE+ SprE+ Kanr Eryr 15
        TX5245TX5241 with pTEX5249; GelE+ SprE+ Kanr Eryr 15
        TX5246TX5242 with pTEX5249; GelE+ SprE+ Kanr Eryr 15
        TX5439TX5264 with pTEX5438; GelE+ SprE+ EryrThis study
        JH2-2bWell-characterized laboratory strain; contains gelE and sprE, lacks fsrAB, and is GelE 5
        TX5440JH2-2 (pTEX5438); GelE+ EryrThis study
        TX5441JH2-2 (pTEX5249); GelE+ EryrThis study
        TX1322bFecal isolate; contains gelE and sprE, lacks fsrAB, and is GelE
        TX5442TX1322 (pTEX5438); GelE+ EryrThis study
        TX5443TX1322 (pTEX5429); GelE+ EryrThis study
Plasmids
    pAT18Shuttle vector derived from pUC18; Eryr 25
    pTEX52496-kb PstI/BglII fragment containing fsrA, fsrB, fsrC, and part of gelE cloned into the shuttle vector pAT18; Eryr 15
    pTEX54381,767-kb PCR product containing gelE and its promoter cloned into pAT18; EryrThis study
  • a Ery, erythromycin; FA, fusidic acid; GelE, gelatinase; SprE, serine protease; S-SprE, superactive serine protease; Kan, kanamycin; Rif, rifampin; A gelatinase phenotype is based on a standard plate assay after 24 h of incubation (15).

  • b JH2-2 and OG1RF both lack asa and esp genes, while TX1322 possesses asa and esp genes.