TABLE 1.

Oligonucleotide primers used in this study

Primer	Nucleotide sequence (5′ to 3′)a	Location	Application	Amplicon
SSP-917B	AAC TGT ACC ACT AAT AAC TCA CAA T	Tn917-lac	PCR
PBSSK3	GTA AAA CGA CGG CCA GT	pBluescript	PCR
Spec1	CGA CGC GTC GAA ATC TAT AAA TAA ACT A	pSF152	Mutagenesis	Spectinomycin cassette
Spec2	CGG TCG ACC GAA ATA ATA AAA CAA AAA A	pSF152	Mutagenesis	Spectinomycin cassette
Fru repressor5′	CGA CGC GTC GGG TAC TAC AAA TGA GTT G	fruR	RT-PCR	fruR-fruI (3,330 bp)
FruPTS3′	CGA CGC GTC GTT ATT YTG ACA ATG GTT T	fruI	RT-PCR	fruR-fruI (3,330 bp)
FruR1	CGG GTA CCC GTC AAT ATC AAC TAC TGA AT	Hypothetical protein 1	Mutagenesis of fruR, RT-PCR	fruR 5′ flanking region (907 bp)
FruR2	CGA CGC GTC GTA TGA ACC AAA CTA TCT A	fruR	Mutagenesis of fruR, RT-PCR	fruR 5′ flanking region (907 bp)
FruR3	CGG TCG ACC GTG TTA AAG TAG CTC CTC T	fruR	Mutagenesis of fruR	fruR 3′ flanking region (740 bp)
FruR4	CGC TCG AGC GTA AGA TTT GAC GAG CAT A	fruK	Mutagenesis of fruR	fruR 3′ flanking region (740 bp)
FruK1	CGG GTA CCC GAC AGG AGT AAA AAC AGA G	fruR	Mutagenesis of fruK	fruK 5′ flanking region (1,022 bp)
FruK2	CGA CGC GTC GGG TTT AAC GTT ACA GTA T	fruK	Mutagenesis of fruK	fruK 5′ flanking region (1,022 bp)
FruK3	CGG TCG ACC GTA TTA AAG AAA CAT ATG A	fruK	Mutagenesis of fruK	fruK 3′ flanking region (834 bp)
FruK4	CGC TCG AGC GTC AGT TCT TCT GTC TTA C	fruI	Mutagenesis of fruK	fruK 3′ flanking region (834 bp)
PTS1	CGG GTA CCC GTT GCT AAG GCA AAG ATA A	fruK	Mutagenesis of fruI	fruI 5′ flanking region (1,053 bp)
PTS2	CGA CGC GTC GCT AGC AAC ATA ACG TCT T	fruI	Mutagenesis of fruI	fruI 5′ flanking region (1,053 bp)
PTS3	CGG TCG ACC GTT TGG TTG GTG CAG TAG T	fruI	Mutagenesis of fruI	fruI 3′ flanking region (613 bp)
PTS4	CGC TCG AGC GCA TTT AGA ACT CCA AAT TAA	Hypothetical protein 2	Mutagenesis of fruI	frul 3′ flanking region (613 bp)
Hypo2rev	ATT CGA CCT TGA ATA TGG TTA ACT TGA G	Hypothetical protein 2	RT-PCR
Hypo3rev	CTC CAA GAT GTG TTC CAA TAA CAC TAC C	Hypothetical protein 3	RT-PCR

^aEngineered 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.