TABLE 1.

Oligonucleotide primers used in this study

PrimerNucleotide sequence (5′ to 3′)aLocationApplicationAmplicon
SSP-917BAAC TGT ACC ACT AAT AAC TCA CAA TTn917-lacPCR
PBSSK3GTA AAA CGA CGG CCA GTpBluescriptPCR
Spec1CGA CGC GTC GAA ATC TAT AAA TAA ACT ApSF152MutagenesisSpectinomycin cassette
Spec2CGG TCG ACC GAA ATA ATA AAA CAA AAA ApSF152MutagenesisSpectinomycin cassette
Fru repressor5′CGA CGC GTC GGG TAC TAC AAA TGA GTT GfruRRT-PCRfruR-fruI (3,330 bp)
FruPTS3′CGA CGC GTC GTT ATT YTG ACA ATG GTT TfruIRT-PCRfruR-fruI (3,330 bp)
FruR1CGG GTA CCC GTC AAT ATC AAC TAC TGA ATHypothetical protein 1Mutagenesis of fruR, RT-PCRfruR 5′ flanking region (907 bp)
FruR2CGA CGC GTC GTA TGA ACC AAA CTA TCT AfruRMutagenesis of fruR, RT-PCRfruR 5′ flanking region (907 bp)
FruR3CGG TCG ACC GTG TTA AAG TAG CTC CTC TfruRMutagenesis of fruRfruR 3′ flanking region (740 bp)
FruR4CGC TCG AGC GTA AGA TTT GAC GAG CAT AfruKMutagenesis of fruRfruR 3′ flanking region (740 bp)
FruK1CGG GTA CCC GAC AGG AGT AAA AAC AGA GfruRMutagenesis of fruKfruK 5′ flanking region (1,022 bp)
FruK2CGA CGC GTC GGG TTT AAC GTT ACA GTA TfruKMutagenesis of fruKfruK 5′ flanking region (1,022 bp)
FruK3CGG TCG ACC GTA TTA AAG AAA CAT ATG AfruKMutagenesis of fruKfruK 3′ flanking region (834 bp)
FruK4CGC TCG AGC GTC AGT TCT TCT GTC TTA CfruIMutagenesis of fruKfruK 3′ flanking region (834 bp)
PTS1CGG GTA CCC GTT GCT AAG GCA AAG ATA AfruKMutagenesis of fruIfruI 5′ flanking region (1,053 bp)
PTS2CGA CGC GTC GCT AGC AAC ATA ACG TCT TfruIMutagenesis of fruIfruI 5′ flanking region (1,053 bp)
PTS3CGG TCG ACC GTT TGG TTG GTG CAG TAG TfruIMutagenesis of fruIfruI 3′ flanking region (613 bp)
PTS4CGC TCG AGC GCA TTT AGA ACT CCA AAT TAAHypothetical protein 2Mutagenesis of fruIfrul 3′ flanking region (613 bp)
Hypo2revATT CGA CCT TGA ATA TGG TTA ACT TGA GHypothetical protein 2RT-PCR
Hypo3revCTC CAA GAT GTG TTC CAA TAA CAC TAC CHypothetical protein 3RT-PCR
  • a Engineered restriction sites are indicated by boldface type. The restriction endonuclease recognition sequences are as follows: KpnI, GGT/ACC; MluI, ACG/CGT; SalI, GTC/GAC; and XhoI, CTC/GAG.