TABLE 2.

PCR primers

PrimerRestriction siteSequencea
Primers used for allelic gene replacement
    ermR2SacI5′-CTC GAG CTC TGA CGG TGA CAT CTC TCT ATT G-3′
    ermL1NheI5′-CTC GCT AGC GAA AAG TAC CAT AAA CGG TCG-3′
    vrsbUR1BamHI5′-CTC GGA TCC AGC GAA AAT ACC AAC CCA CG-3′
    vrsbUL1SacI5′-CTC GAG CTC GAA ATG CGC CTC CTT ACT TC-3′
    hrsbUR1NheI5′-CTC GCT AGC GAT GGT GTT ACA GAG GCA CG-3′
    hrsbUL1KpnI5′-CTC GGT ACC AGC TGG CAA CCG CAT TTC-3′
    hrsbWR1NheI5′-CTC GCT AGC GTT ATT TCA GAC CAA GGT G-3′
    hrsbWL1KpnI5′-CTC GGT ACC TTA TCA TTC TGT TGT CCC AT-3′
    vsigBR1BamHI5′-CTC GGA TCC CCA ATG AGA CAA GAA GGC AC-3′
    vsigBL1SacI5′-CTC GAG CTC CTT GAG CTT GGC TAT CTT CG-3′
    hsigBR1NheI5′-CTC GCT AGC CGA AAG AAG CTC AGG TGG AC-3′
    hsigBL1KpnI5′-CTC GGT ACC GAT GCT GAA TAA ACT GAT GCG-3′
Primers used for Northern blot hybridization probes
    erm.forNone5′-AAT TGG AAC AGG TAA AGG GC-3′
    erm.revNone5′-AAC ATC TTG GGT ATG GCG G-3′
    sigB.forNone5′-AGA TTT AGT TCA AGT TGG TA-3′
    sigB.revNone5′-TTA TCA TCT TGT TGT CCC AT-3′
    rsbU.forNone5′-GAA GTG GAA GTA AGG AGG CG-3′
    rsbU.revNone5′-TCG ATG TGT TAC CAG AAG TCG-3′
    sarA.forNone5′-ATA GGG AGG TTT CAT TAA TGG C-3′
    sarA.revNone5′-TTT GCT TCT GTG ATA CGG TTG-3′
    asp23.forNone5′-AAA ATC AAA AAG CAC TTG AGC G-3′
    asp23.revNone5′-AAA AAA TTG CAG GTAT TGC AG-3′
    icaA.forNone5′-GAA TCC AAA ATT AGG CGC AG-3′
    icaA.revNone5′-AAC ATC CAG CAT AGA GCA CG-3′
    icaR.forNone5′-TCC GAA AAG GGG TAC GAT G-3′
    icaR.revNone5′-CCT CTT TAT CCA AAG CGA TG-3′
Primers used for quantitative RT-PCR
    asp23.real1None5′-TCC AAC TTC TAC AGA TAC GCC-3′
    asp23.real2None5′-AAA ATT GCA GGT ATT GCA GC-3′
    icaA.real1None5′-TGT ATC AAG CGA AGT CAA TCT C-3′
    icaA.real2None5′-GGC ACT AAC ATC CAG CAT AG-3′
    icaR.real1None5′-TGA AGA TGG TGT TTG ATT TGT G-3′
    icaR.real2None5′-CCA TTG ACG GAC TTT ACC AG-3′
    rsbU.real1None5′-AGC GTT TGA GGA AAT TGG TGT G-3′
    rsbU.real2None5′-CCT CTA CAT CTC GTG CCT CTG-3′
    gyrB.real1None5′-CTG ACA ATG GCC GTG GTA TTC-3′
    gyrB.real2None5′-GAA GAT CCA ACA CCG TGA AGA C-3′
  • a Restriction sites are underlined.