Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Characteristicsa | Reference or source |
|---|---|---|
| Strains | ||
| S. flexneri | ||
| PE638 | S. flexneri Y rpoB (Rifr) | 14 |
| RMM109 | PE638 Δwzy; Rifr | 3 |
| RMA4337 | RMM109 Δwzz (Rifr Tetr) | This study |
| PNRM6 | RMM109(pAC/pBADT7-1) | This study |
| PNRM11 | PNRM6(pWaldo-TEV-GFP) | This study |
| PNRM13 | PNRM6(pRMPN1) | This study |
| PNRM75 | PNRM6(pRMPN7) | This study |
| PNRM76 | PNRM6(pRMPN8) | This study |
| PNRM77 | PNRM6(pRMPN9) | This study |
| PNRM78 | PNRM6(pRMPN10) | This study |
| PNRM79 | PNRM6(pRMPN11) | This study |
| PNRM80 | PNRM6(pRMPN12) | This study |
| PNRM81 | PNRM6(pRMPN13) | This study |
| PNRM82 | PNRM6(pRMPN14) | This study |
| PNRM83 | PNRM6(pRMPN15) | This study |
| PNRM84 | PNRM6(pRMPN16) | This study |
| PNRM85 | PNRM6(pRMPN17) | This study |
| PNRM119 | PNRM6(pRMPN19) | This study |
| PNRM120 | PNRM6(pRMPN21) | This study |
| PNRM121 | PNRM6(pRMPN22) | This study |
| PNRM122 | PNRM6(pRMPN23) | This study |
| PNRM123 | PNRM6(pRMPN24) | This study |
| PNRM124 | PNRM6(pRMPN25) | This study |
| PNRM126 | RMA4337(pAC/pBADT7-1) | This study |
| PNRM134 | PNRM126(pRMPN1) | This study |
| PNRM131 | PNRM126(pRMPN7) | This study |
| PNRM132 | PNRM126(pRMPN15) | This study |
| PNRM133 | PNRM126(pRMPN16) | This study |
| PNRM136 | PNRM126(pRMPN8) | This study |
| PNRM137 | PNRM126(pRMPN10) | This study |
| PNRM140 | PNRM126(pRMPN13) | This study |
| PNRM141 | PNRM126(pRMPN14) | This study |
| PNRM142 | PNRM126(pRMPN9) | This study |
| PNRM143 | PNRM126(pRMPN11) | This study |
| PNRM144 | PNRM126(pRMPN19) | This study |
| PNRM145 | PNRM126(pRMPN24) | This study |
| PNRM146 | PNRM126(pRMPN25) | This study |
| PNRM147 | PNRM126(pRMPN23) | This study |
| PNRM148 | PNRM126(pRMPN21) | This study |
| PNRM149 | PNRM126(pRMPN22) | This study |
| PNRM150 | PNRM126(pRMPN12) | This study |
| PNRM151 | PNRM126(pRMPN17) | This study |
| E. coli | ||
| XL10G | Tetr Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tetr) Cmr] | Stratagene |
| Lemo21(DE3) | fhuA2 (lon) ompT gal (λ DE3) (dcm) ΔhsdS/pLemo (Cmr) | New England Biolabs |
| PNRM15 | Lemo21(DE3)(pRMPN1) | This study |
| Plasmids | ||
| pRMCD6 | Source of wzySf (modified codons at positions 4, 9, and 23) | 18 |
| pAC/pBADT7-1 | Source of T7 RNA polymerase; Cmr | 34 |
| pWaldo-TEV-GFP | Cloning vector with GFP tag; Kmr | 28 |
| pRMPN1 | pWaldo-wzySf-GFP; Kmr | This study |
| pRMPN7 | pRMPN1 with G130V point mutation in the wzySf gene | This study |
| pRMPN8 | pRMPN1 with L111I point mutation in the wzySf gene | This study |
| pRMPN9 | pRMPN1 with N86K point mutation in the wzySf gene | This study |
| pRMPN10 | pRMPN1 with L28V point mutation in the wzySf gene | This study |
| pRMPN11 | pRMPN1 with P165S point mutation in the wzySf gene | This study |
| pRMPN12 | pRMPN1 with G82C point mutation in the wzySf gene | This study |
| pRMPN13 | pRMPN1 with N147K point mutation in the wzySf gene | This study |
| pRMPN14 | pRMPN1 with L191F point mutation in the wzySf gene | This study |
| pRMPN15 | pRMPN1 with L214I point mutation in the wzySf gene | This study |
| pRMPN16 | pRMPN1 with P352H point mutation in the wzySf gene | This study |
| pRMPN17 | pRMPN1 with V92 M point mutation in the wzySf gene | This study |
| pRMPN19 | pRMPN1 with F52Y point mutation in the wzySf gene | This study |
| pRMPN21 | pRMPN1 with F52C/I242T point mutations in the wzySf gene | This study |
| pRMPN22 | pRMPN1 with C60F point mutation in the wzySf gene | This study |
| pRMPN23 | pRMPN1 with Y137H point mutation in the wzySf gene | This study |
| pRMPN24 | pRMPN1 with L49F/T328A point mutation in the wzySf gene | This study |
| pRMPN25 | pRMPN1 with F54C point mutation in the wzySf gene | This study |
| pCACTUS | Suicide vector containing sacB, Cmr gene, and orits | 14 |
| pRMA577 | Suicide vector containing SphI-SphI fragment with the rol gene | 14 |
| pCACTUS-wzzSf::Tcr | Suicide mutagenesis construct to construct the strain RMA4337 | This study |
a
Rifr, rifampin resistant; Kmr, kanamycin resistant; Cmr, chloramphenicol resistant; Tetr, tetracycline resistant. The λ DE3 genotype is λ sBamHIo ΔEcoRI-B int::(lacI::PlacUV5::T7 gene 1) i21.