Table 1.

Strains and plasmids used in this work

Strain or plasmidRelevant characteristicsaReference or source
E. coli
 DH5αFrm+ø80dlacZΔM15Gibco BRL
 MC1061FaraD139Δ(ara-leu)7696 galE15 galK16Δ(lac)X74 rpsL (Strr)hsdR2 (rkmK+) mcrA mcrBl41
 BL21 (DE3)Expression host, inducible T7 RNA polymeraseNovagen
Streptococcus
 RH110E. faecalisOG1RF::Tn916ΔE rif fus erm28
 12021S. mitisThis study
 SF100Smr derivative of S. mitisstrain 12021This study
 PS101SF100pblT::Tn916ΔEThis study
 PS116SF100pblA::Tn916ΔEThis study
 PS163SF100 pblT::pVA891This study
 PS301SF100 pblA::pVA891This study
 PS344SF100Δ[ORF1-pblB]This study
 PS345SF100 pblB::pVA891This study
 PS361SF100 pblR::pVA891This study
 PS385PS301(pDC123)This study
 PS386PS301(pSF116A)This study
 PS389PS345(pDC123)This study
 PS390PS345(pSF116B)This study
 PS416PS301(pSF116B)This study
Plasmids
 pWE15Cos, Apr39
 pBluescriptAprStratagene
 pVA891Tcr, cat, ori (E. coli), erm (gram positive)18
 pDC123cat phoZ, streptococcal expression vector with blue-white screening7
 pSF116ApblA cloned in pDC123This study
 PSF116BpblB cloned in pDC123This study
  • a Smr, streptomycin resistance; Apr, ampicillin resistance; Tcr, tetracycline resistance. Cos, recognition site for packaging by phage lambda.