TABLE 2.

Oligonucleotides used in this study

Function and name	Sequencea	Locationb	Utilization
Construction of mutants
gutbc1	CTGCCATACCAGCAATGGC	−285	Cloning gutB with an internal 480-bp deletion
gutbc2	TGCCAGCGGACCAGACTTC	+315
gutbc3	TATCGTCATCGGCTTTATCTG	+796
gutbc4	ATCCTGCGAGCTCAGGTG	+1455
gutF1	GCAAGTAAAGGCATC	−350	Cloning part of gutR to create a frameshift at
gutR1	TGGTGAATCGTCCATCAAG	+689	PstI restriction site (codon 48)
gutR2	TGATGCTGGCAGAATCATC	−451	Cloning gutM to create a frameshift at PstI
gutC1	TGGAACAGCTTCATGAAGC	+574	restriction site (codon 16)
Complementation of mutants
gutRBglII	CATAAGATCTGGGAGGTAACCTG	−24	Cloning gutR in pT1NX
gutRSpeI	TATCAACTAGTTCATTTAAAGTCCCTC	+1884
gutMBglII	CCTTAACAGATCTCACGGAGGGAC	−29	Cloning gutM in pT1NX
gutMSpeI	CATCTACTAGTTTTAAATTGTATTC	+522
Transcription analysis
ribu1	TATTATGGGCAGGATAATGC	−302	DNA probe (the primer location is indicated
gutF3	GCCGTTAACAACCGTAGCG	+117	with respect to guF)
gut3	GACTTTAAGGACCCTAA	+133	RT-PCR analysis (the primer location is
gutbc4	ATCCTGCGAGCTCAGGTG	+5277	indicated with respect to guF)
gut5	TTTTCAACTTCGTCAGGATC	+197	Identification of the transcription start site of
gutF3	GCCGTTAACAACCGTAGCG	+117	the gut operon (the primer location is indicated with respect to guF)
Overexpression of genes
gutRBamHI	GGGAGGGATCCGGATTGAATAG	−14	Cloning gutR in pQE30
gutRSalI	CAAAACGTCGACTTAAAGTCCC	+1881
gutMBamHI	CGGAGGGATCCAAAATGAACG	−14	Cloning gutM in pQE30
gutMHindIII	CTCCTAAGCTTAAATTGTATTC	+519
Gel mobility shift and DNase I footprinting
gut8D	GTCAAGCACCGCTAACC	−200	Amplified 179-bp DNA fragment of the gut
gutF4	TTTCTGCTCTATGGTAGC	−22	promoter region (the primer location is indicated with respect to guF)

^aLetters in italics indicate newly created restriction sites.

^bThe numbers indicate the position of the 5′ end of the primer with respect to the start codon of the gene specified in the “Utilization” column.