Table 1

Bacterial strains and plasmids used in this work
Bacterial strains or plasmid Characteristics Source or reference
Shigella flexneri
S. flexneri 2457T Wild type strain Laboratory stock
S. flexneri 2457T ΔgluQ-rs::kan Deletion mutant of gluQ-rs gene This work
Escherichia coli
E. coli W3110 ΔgluQ-rs::kan Deletion mutant of gluQ-rs gene [10]
DH5α F-ϕ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK-, mK+) phoA supE44 λ-thi-1 gyrA relA1 [24]
BL21(DE3) F-ompT gal dcm lon hsdSB(rB-mB-) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) Invitrogen
Plasmids
pTZ57R/T bla, pMB1 ori, lacZ peptide, f1 phage ori Fermentas®
pQF50 bla, pMB1 ori, lacZ gene without promoter [23]
pET15c Empty vector, a modified version of pET15b This work
pVCDT S. flexneri fragment from nucleotide +58a of dksA gene to beginning of gluQ-rs gene (+590) cloned into pQF50. Pair of primers used were PgluQF/PdksARCT. This work
pVCPDT S. flexneri fragment from nucleotide −506 of dksA gene to beginning of gluQ-rs gene (+590) cloned into pQF50. Pair of primers used were PdksAF/PdksARCT. This work
pVCPDTMut S. flexneri fragment from nucleotide −506 of dksA gene to beginning of gluQ-rs gene (+590) cloned into pQF50, with the terminator mutated by the nucleotides indicated in Figure 4a. This work
pVCPD S. flexneri fragment from nucleotide −506 of dksA gene to nucleotide +527 (end of dksA gene) cloned into pQF50. Pair of primers used were PdksAF/PdksARST. This work
pATGGQRS S. flexneri gene from nucleotide +509 (stop codon of dksA) to nucleotide +1469 (last codon of gluQ-rs without stop codon). Pair of primers used were ATGGQRSF/ATGGQRSR. This work

aFragments cloned are numbered based on the transcription start of dksA identified by [25].

Caballero et al.

Caballero et al. BMC Microbiology 2012 12:226   doi:10.1186/1471-2180-12-226

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