TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or feature(s)a | Reference or source |
|---|---|---|
| Strains | ||
| UTI89 | Clinical cystitis isolate | 44 |
| UTI89 attλ::PSSH10-1 | Promoterless gfp; spectinomycin resistance cassette at lambda integration site | 73 |
| MG1655 | K-12 reference strain | 8 |
| MG1655 attλ::PSSH10-1 | Promoterless gfp; spectinomycin resistance cassette at lambda integration site | 73 |
| UTI89 kpsF:: EP185 | Insertion of pEP185.2 in kpsF; polar inactivation of region I capsule genes | This study |
| UTI89 Δkps | ΔkpsFEDUCS::FRTΨ | This study |
| MG1655 attHK022::COM-GFP | tac-gfp in single copy at the HK022 phage integration site, Kanr | 73 |
| UTI89 Δkps attHK022::COM-GFP | ΔkpsFEDUCS::FRTΨtac-gfp; kanamycin resistance cassette | This study |
| MG1655 attλ::kps-gfp | PRegI::gfp at lambda integration site; Specr | This study |
| UTI89 attλ::kps-gfp | PRegI::gfp at lambda integration site; Specr | This study |
| UTI89 Δneu | ΔneuDBACES::FRTΨ | This study |
| UTI89 ΔcdiA | ΔcdiA::kan | This study |
| UTI89 Δkps ΔcdiA | ΔkpsFEDUCS::FRTΨ ΔcdiA::kan | This study |
| JW3195.1 | ΔnanR::kan | 5 |
| UTI89 Δkps ΔnanR | ΔkpsFEDUCS::FRTΨ ΔnanR::kan | This study |
| Plasmids | ||
| pPSSH10 | Promoterless gfp, Specr | 73 |
| pINT | Lambda integrase, Ampr | 49 |
| pCOM-GFP | tac-gfp, Ampr, Kanr | 66 |
| pKD4 | FRT-Kan-FRT, Ampr | 18 |
| pCP20 | FLP, Ampr | 13 |
| pEP185.2 | SacB, Cmr | 19 |
| pBR329 | Ampr, Cmr, Tetr | 15 |
| pSX50 | kpsFEDUCS, Ampr, Cmr | 61 |
| pGA122 | kps-gfp in pPSSH10-1 | This study |
| pBAC-LacZ | Mini F′ single-copy plasmid, Cmr | Addgene plasmid 13422 |
| pBAC-LacZ Reg I kps | EcoRI fragment of pSX50 containing kpsFEDUCS, Cmr | This study |
| pCR-nanR | nanR from UTI89 in pCR-BLUNT II, Kanr | This study |
| pBAC-LacZ nanR | nanR from pCR-nanR in pBAC-LAC, Cmr | This study |
a
Superscript Ψ Indicates residual interrupting scar after removal of antibiotic cassette by FLP recombinase.