Table 1.
Bacterial strains and plasmidsa
| Strain or plasmid | Relevant genotype, phenotype, or descriptionb | Source or referencec |
|---|---|---|
| Strains | ||
| DH5α | For general DNA manipulations and detection of bacteriophage | Stratagene |
| TG1 | For general DNA manipulations and detection of bacteriophage | Stratagene |
| SM10λpir | pir traRP4 | 33, IAH |
| WS-2773E | O141:H5 CS5 CS6 LT ST EAST1 (clinical isolate) | NMRC |
| ACAM2006 | O141:H5 CS5 CS6 ΔastA ΔeltAB ΔestA ΔompC ΔompF ΔaroC | This work |
| ACAM2012 | O141:H5 CS5 CS6 ΔastA ΔeltAB ΔestA ΔompC ΔompF ΔaroC Δphage | This work |
| ACAM2022 | O141:H5 CS5 CS6 ΔastA ΔeltAB ΔestA ΔompF::Ptac-LTB ΔaroC Δphage | This work |
| WS-3504D | O39:H12 CS2 CS3 LT ST EAST1 Apr (clinical isolate) | NMRC |
| ACAM2007 | O39:H12 CS2 CS3 ΔastAΔeltAB ΔestA ΔompC ΔompF ΔaroC | This work |
| ACAM2017 | O39:H12 CS2 CS3 ΔastA ΔeltAB ΔestA ΔompC::CS1 ΔompF ΔaroC | This work |
| ACAM2027 | O39:H12 CS2 CS3 ΔastA ΔeltAB ΔestA ΔompC::CS1 ΔompF::Ptac-LTB ΔaroC | This work |
| E1392/75 | O6:H16 CS1 CS3 ST LT Sur Smr | 5 |
| E1392/75-2A | O6:H16 CS1 CS3 Sur Smr | 5 |
| PTL003 | O6:H16 CS1 CS3 Sur Smr ΔaroC ΔompC ΔompF | 32 |
| ACAM2008 | O6:H16 CS1 CS3 ΔaroC ΔompC ΔompF | This work |
| WS-1858B | O71:H− CFA/I ST EAST1 Apr Tpr Sur (clinical isolate) | NMRC, 31 |
| ACAM2010 | O71:H− CFA/I ΔastA ΔestA ΔaroC ΔompC ΔompF | 31 |
| ACAM2025 | O71:H− CFA/I ΔastA ΔestA ΔaroC ΔompC ΔompF::pLLTB | This work |
| Plasmids | ||
| RP4 | 60-kb plasmid, covalently closed circular DNA size standard | 31 |
| pvirF98 | ∼90-kb plasmid from Salmonella enterica serovar Typhimurium F98, covalently closed circular DNA size standard | 31 |
| ColVIk94 | ∼127-kb plasmid, covalently closed circular DNA size standard | 31 |
| pKK223-3 | Expression vector, source of Ptac and rrnB transcription terminator | Amersham-Pharmacia |
| pPCR-Script Amp SK+ | Plasmid vector | Stratagene |
| pPCRProm | Native LT promoter in pPCR-Script | This work |
| pACYC184 | Plasmid vector | 3 |
| pACYC-Tc | ori15A Tcr | 31 |
| pACYC-CS1 | CS1 operon in pACYC184 | This work |
| pACYC-csaA | CS1 csaA gene in pACYC184 | This work |
| pACYC-PtacLTB | Ptac-LTB construct in pACYC184 | This work |
| pACYC-PtacLTBrrnB | Ptac-LTB-rrnB transcription terminator construct in pACYC184 | This work |
| pACYC-ΔompC | This work | |
| pACYC-ΔompC::cooBA | This work | |
| pJCB12 | oriR6K mobRP4 sacB cat | 31 |
| pJCB12-ΔeltA | 658-bp deletion construct of LTA gene in pJCB12 | This work |
| pJCB12- ΔestAE | ST gene deletion fragment in pJCB12 | This work |
| pJCB12-ΔastA | 286-bp fragment with 42-bp defined deletion in EAST1 gene in pJCB12 | 31 |
| pJCB12-ΔompC | 1,016-bp fragment with deletion of the whole ompC gene in pJCB12 | 31 |
| pJCB12-ΔompF | 943-bp fragment with deletion of the whole ompF gene in pJCB12 | 31 |
| pJCB12-ΔompF::LTB | This work | |
| pJCB12-ΔompF::PtacLTB | This work | |
| pJCB12-ΔompF::PtacLTBrrnB | LTB expressed from tac promoter, incorporating rrnB transcriptional terminator and flanked by ΔompF nucleotide sequences in pJCB12 | This work |
| pJCB12-ΔompF::LLTB | This work | |
| pJCB12-ΔompC::CS1 | CS1 operon flanked by ΔompC nucleotide sequences in pJCB12 | This work |
| pJCB12-ori15A | pACYC184 replication origin in pJCB12 | 31 |
| pTETnir15 | Inducible expression vector | 4 |
| pLLTB | pTETnir15 expressing LTB from its native promoter | This work |
a
For the sake of brevity, some strains are not listed.
b
Antibiotic resistance: Nalr, nalidixic acid; Apr, ampicillin; Tpr, trimethoprim; Smr, streptomycin; Sur, sulfamethoxazole; Tcr, tetracycline.
c
IAH, provided by P. Barrow, Institute for Animal Health, Compton, United Kingdom; NMRC, provided by S. Savarino, Naval Medical Research Center, Silver Spring, MD.