Bacterial strains or plasmid | Characteristics | Source or reference |
---|---|---|
Shigella flexneri | Â | Â |
S. flexneri 2457T | Wild type strain | Laboratory stock |
S. flexneri 2457T ΔgluQ-rs::kan | Deletion mutant of gluQ-rs gene | This work |
Escherichia coli | Â | Â |
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 hsdS B (r B - m B - ) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) | Invitrogen |
Plasmids | Â | Â |
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 |