Table 2

Oligonucleotides used in this study

NameSequence (5′–3′)aPurpose
relA-1-fwCCCCTCGAGGAAGGCGCGCGGACAAGCGGMutagenesis
relA-2-rvCGGCGCTCACTGCAGCATCGTCGGGTATGGCCAGTTMutagenesis
relA-3-fwCCGACGATGCTGCAGTGAGCGCCGCCGGTGGGCCGCMutagenesis
relA-4-rvCCCGCATGCCGTATCGACGAGTTCGCCGTCMutagenesis
spoT-1-fwGCGGTCGACCTGCCGCCGTCGCTCGCGGCMutagenesis
spoT-2-rvGCGGCGCTAACTAGTCATTTTCGCCTCCTGGGTTCGMutagenesis
spoT-3-fwGCGAAAATGACTAGTTAGCGCCGCGCGGCGCCCGACMutagenesis
spoT-4-rvGCGCCCGGGGCCTGCGCGAAATCGAGGTCGMutagenesis
relA-up-fwACAACGGCACCGTCTATCTCConfirmation of plasmids
relA-down-rvGCGGTGTCATAACCGTATTCConfirmation of plasmids
spoT-up-fwCTCGCATTACCGTTGAAGACConfirmation of plasmids
spoT-down-rvGCCCTTTCAGTTCGAAGTTGConfirmation of plasmids
relA-3-fwGCGTGCAGTCGGACCTGCTGConfirmation of mutation
relA-4-rvGTTCGGGAAAATTCGCAAGCCConfirmation of mutation
relA-5-wtGTGGAAGCGGAATCGGCTGConfirmation of mutation
spoT-3-fwGGTGAAGAAGCAGTTCCGCAConfirmation of mutation
spoT-4-rvCGCCCGAAACAAATGGCGGConfirmation of mutation
spoT-5-wtGCGGACGATGGTGTAGTGCConfirmation of mutation
relA-RT-1AGATCCGCACGCAGGAAATGRT-PCR analysis
relA-RT-2TTCGCTGTGCAGGTGGTAAGRT-PCR analysis
spoT-RT-1GCGCATCAATGGCTCAAGTCRT-PCR analysis
spoT-RT-2TTCAGGCGCATCGTCTTCAGRT-PCR analysis
16S-RT-1GCCAGTCACCAATGCAGTTCRT-PCR normalization
16S-RT-2ACCAAGGCGACGATCAGTAGRT-PCR normalization
  • a Restriction sites are underlined. Complementary regions are in italics. Translational start and stop codons are in bold.