TABLE 1.
Shigella strains and plasmids used in this study
| Strain or plasmid | Relevant genotype and/or phenotype or feature | Source or reference |
|---|---|---|
| Strains | ||
| 2457T | Wild-type S. flexneri serotype 2a | 18 |
| BS103 | Virulence plasmid-cured 2457T | 35 |
| BS260 | 2457T mxiA::lacZ | 6 |
| BS473 | 2457T streptomycin resistant | Lab stock |
| BS547 | 2457T mxiM1 (mxiM::aphA-3) | This work |
| BS548 | BS547/pRRS4 (mxiM1PBAD-mxiM+) | This work |
| BS575 | BS547/pRRS5 (mxiM1/PLAC-mxiM+) | This work |
| BS576 | BS547/pBAD18 | This work |
| BS588 | BS547/pBluescript SK(+) | This work |
| BS599 | 2457T/pRRS12 (mixiM+/PBAD-mxiM2) | This work |
| BS603 | BS547/pRRS12 (mxiM1/PBAD-mxiM2) | This work |
| Plasmids | ||
| pBAD18 | Arabinose-inducible PBAD expression vector | 23 |
| pET19b | Vector used to construct histidine-tagged MxiM | Novagen |
| pGP704 | Suicide vector used to disrupt mxiM | 41 |
| pRRS1 | 1850-bp PCR generated fragment, extending from 912 bp upstream of the mxiM start codon to 511 bp downstream of the stop codon, ligated with EcoRI-HindIII-digested pUC19 | This work |
| pRRS2 | 840-bp SmaI fragment of pUC18K (38), bearing aphA-3, ligated in the proper orientation with BspMI-digested (Klenow-treated) pRRS1 | This work |
| pRRS3 | 2690-bp HindIII (Klenow-treated)-EcoRI fragment of pRRS2, bearing the mxiM::aphA-3 allele, ligated with EcoRV-digested pGP704 | This work |
| pRRS4 | 484-bp PCR-generated fragment, extending from 19 bp upstream of the mxiM start codon to 26 bp downstream of the stop codon, ligated with EcoRI-HindIII-digested pBAD18 | This work |
| pRRS5 | 484-bp EcoRI-HindIII fragment of pRRS4, bearing mxiM, ligated with EcoRI-HindIII-digested pBluescript SK(+) | This work |
| pRRS11 | 370-bp PCR-generated fragment, extending from 67 bp downstream of the mxiM start codon (thus deleting signal sequence) to 10 bp downstream of the stop codon, ligated with NdeI-BamHI-digested pET19b | This work |
| pRRS12 | Mutagenized mxiM allele encoding the G23R substitution, ligated with EcoRI-HindIII-digested pBAD18 | This work |