TABLE 4

Primers used in this study

Primer namePurposeSequence (5′–3′)a
agr-F1PCRTTAGCTCTTTATATTGGATATACAG
agr-R1PCR/mutant verification/RT-PCRCCGGTTTAAAACCGACCTTTAG
agrBFMutant verification/RT-PCRGATGTTAGCCATGTATGCTTTCG
agrF1-SacIagrB complementationTAGAGAGCTCTTAGCTCTTTATATTGGATATACAG
agrR1-BamHIagrB complementationTAGAGGATCCCCGGTTTAAAACCGACCTTTAG
CT_netB-F2Mutant verificationAGTGTAATTAGTACAAGCC
qNetB-RMutant verificationGGCCATTTCATTTTTCCGTAA
CTC-netFnetB complementationCGGGATCCGTACCATTTAAATTAAGCAC
CTC-netRnetB complementationTCGAGCTCCCCTCTATATACTATTGATTG
mNetB-FNetB protein expressionCGGCGAATTCAGTGAATTAAATGACATAAACA
mNetB-RNetB protein expressionCGGCAAGCTTCAGATAATATTCTATTTTATGA
virRP-L KpnIMutant verification/virR complementation/RT-PCRTTGGTACCTTATGTTCATAAAATAGAAAGTGG
virR-R XbaIMutant verification/virR complementation/RT-PCRTTTCTAGATTAACATATTAAATCCCCTAAAAGGC
luxS-1FMutant verification/RT-PCRGTGGTTAAAGTTGAAAGTTTG
luxS-1RMutant verification/RT-PCRATATCTTAACATCAATATCTCC
luxS-F3-KpnIluxS complementationTTGGTACCATTAGCTGATCCAGTAGGTTC
luxS-R3-XbaIluxS complementationTTTCTAGACTTTAGATTATAAATAGTAATTAC
ErmFMutant verificationGATATTCACCGAACACTAGG
ErmRMutant verificationTTACCTGTTCCAATTTCGTA
CT_probe-F1Southern blottingACAACTTAATTATACCCACT
CT_probe-R1Southern blottingCTTGTGTTTATGAATCACG
QnetBFqPCRAGTGTAATTAGTACAAGCC
QnetBRqPCRGGCCATTTCATTTTTCCGTAA
rpoA-F1qPCRTTACCTGGAGTGGCTCCAAC
rpoA-R1qPCRACACCTGGTCCTTGAGCATC
CP4_0573-F1qPCRAAGCCATCAGATACGGTGGT
CP4_0573-R1qPCRAGATTCGGCTATCCTTTGCAT
CP4_0575-F1qPCRTCATCATGCAGATGCTTCCT
CP4_0575-R1qPCRCACCTTTTGCATCAGTATTGACA
rSAM-FqPCRGAGTTTTTCCAGATGGTTC
rSAM-RqPCRGATGTAAATTTATGGCTATAC
  • a Restriction enzyme sites are underlined.