TABLE 1.

Bacterial strains, plasmids, and oligonucleotides used in this study

Strain, plasmid, or oligonucleotide	Relevant characteristics or sequencea	Source, reference, or purpose
Strains
E. coli DH5	supE44 hsdR17 recA1 endA1 gyrA96 thi-1 relA1	8
E. coli BL21 (DE)	ompT hsdSB (rB− mB−) gal dcm (DE3)	5
C. glutamicum WT	Wild-type strain ATCC 13032	American Type Culture Collection
C. glutamicum RG1	WT C. glutamicum with truncated ramB gene, shortened by 775 bp	6
C. glutamicum RG2	WT C. glutamicum with truncated ramA gene, shortened by 364 bp	4
Plasmids
pET2	Multicopy promoter probe vector carrying the promoterless cat gene; Kmr	17
pRIM2	Integrative promoter probe vector carrying the promoterless cat gene; Kmr	17
pET28-RamAx6His	pET28 containing the ramA structural gene	4
pET29-RamBx6His	pET29 containing the ramB structural gene	6
pramBp1	pET2 containing the 593-bp ramB promoter fragment	This work
pramBp3	pET2 containing the 370-bp ramB promoter fragment	This work
pramBp3b	pET2 containing the 211-bp ramB promoter fragment	This work
pramBp3c	pET2 containing the 135-bp ramB promoter fragment	This work
pRIM-ramBp1	pRIM containing the 593-bp ramB promoter fragment	This work
pRIM-ramBp3c	pRIM containing the 135-bp ramB promoter fragment	This work
pDrive	Cloning vector; Kmr AmprlacZα orif1 ori-pUC	QIAGEN GmbH, Hilden, Germany
pDrive ramBpRACE	pDrive derivative plasmid containing the ramB PCR-amplified fragment from the RACE assay	This work
Oligonucleotidesb
ramBpLF forw	5′-ACGCGTCGACCTAACAGTCATGGCACCTCCAGTGTGG-3′	ramBp1and ramBp5
ramBpLF rev	5′-CGCGGATCCCAAGGGTTGCTGCTAAGGATGCCTG-3′	ramBp1, ramBp3, ramBp3a, ramBp3b, ramBp3c
ramBp forw	5′-AGCGAAAATCAACAAGTTTGCAACACCTCAGT-3′	ramBp 13-bp motif
ramBp rev	5′-ACTGAGGTGTTGCAAACTTGTTGATTTTCGCT-3′	ramBp 13-bp motif
ramBpKF forw	5′-ACGCGTCGACGCTTCCTCACAGGATACCGA-3′	ramBp3
ramBp3a forw	5′-CAGGGAGCAACTTTGCGCAG-3′	ramBp3a
ramBp3b forw	5′-ACGCGTCGACGATGTGGCCCGACCACGCCG-3′	ramBp3b
ramBp3c forw	5′-ACGCGTCGACCTCAGTGCCAAGAGTGGTTA-3′	ramBp3c
ramBp5 rev	5′-CGCAGGTAGAGCACACTCAAT-3′	ramBp5
CM4	5′-GAAAATCTCGTCGAAGCTCG-3′	cDNA synthesis; 5′-RACE ramA and ramB transcriptional start site determination
Oligonucleotide dT anchor primer	5′-GACCACGCGTATCGATGTCGACTTTTTTTTTTTTTTTTV-3′	Amplification of dA-tailed cDNA (Roche, 5′/3′ RACE kit)

^aRestriction sites in the oligonucleotides are underlined. V represents an A, C, or G.

^bforw., forward; rev, reverse.