TABLE 1.
Bacterial strains and plasmids in this study
| Strain or plasmid | Genotype | Source or reference |
|---|---|---|
| Strains | ||
| EK227 | K-12, wild type, λ− F− | A. C. Matin |
| EK344 | (GE1050) Δcrp::Cm | 16 |
| EK395 | Δcya-1400::Km λ− e14−relA1 spoT1 thi-1 | J. Kaper |
| EK432 | Δ(lacA-lacZ)515(::Cat) | 3 |
| EK426 | K-12 (MG1655) ΔgadX::Km | T. Conway |
| EK437 | hns-206oppC506::Tn10 | C. Higgens |
| EK441 | K-12 (MG1655) ΔgadW::Km | T. Conway |
| EK442 | K-12 (MG1655) ΔgadXW::Km | T. Conway |
| EK540 | TOPO 10 F−mcrA Δ(mrr-hsdRMS-merBC) φ80 lacZΔM15 ΔlacX74 recA1 deoR araD139 Δ(ara-leu)7697 galU galK rpsL endA1 nupG | Invitrogen |
| EK541 | BL21 Star(DE3): F−ompT hsdB (rB− mB−) dcm rne-131 | Invitrogen |
| EF362 | K-12 (EK227) rpoS::Tn10 | 2 |
| EF528 | K-12 (EK227) Δcrp::Cm | EK227 × EK344 |
| EF615 | EK298 trpDC::putPA1303-Km-gadA::lacZ(o) (−165 to +788) | 2 |
| EF646 | Δcya-1400::Km/pT18-Zip × pT25-Zip | 14 |
| EF663 | EK298 trpDC::putPA1303-Km-gadA::lacZ(o) (−51 to +788) | 2 |
| EF666 | EK298 trpDC::putPA1303-Km-gadA::lacZ(o) (−86 to +788) | 2 |
| EF678 | K-12 (EK227) Δcrp::Cm rpoS::Tn10 | EF528 × EF362 |
| EF757 | K-12 (EK227) ΔgadX::Km | EK227 × EF426 |
| EF825 | K-12 (EK227) Δ(lacA-lacZ)::Cat | EK227 × EK432 |
| EF828 | K-12 (EK227) ΔgadX | EF757 × pCP20 |
| EF830 | K-12 (EK227) ΔgadX Δ(lacA-lacZ)::Cat | EF828 × EK432 |
| EF833 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−165 to +788) Δ(lacA-lacZ)::Cat | EF825 × EF615 |
| EF839 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−165 to +788) ΔgadX Δ(lacA-lacZ)::Cat | EF830 × EF615 |
| EF861 | K-12 (EK227) ΔgadW::Km | EK227 × EK441 |
| EF863 | K-12 (EK227) ΔgadXW::Km | EK227 × EK442 |
| EF864 | K-12 (EK227) ΔgadW::Km Δcrp::Cm | EF528 × EK441 |
| EF865 | K-12 (EK227) ΔgadXW::Km Δcrp::Cm | EF528 × EK442 |
| EF906 | K-12 (EK227) ΔgadX Δcrp::Cm | EF828 × EF528 |
| EF907 | K-12 (EK227) ΔgadW | EF861 × pCP20 |
| EF908 | K-12 (EK227) ΔgadXW | EF863 × pCP20 |
| EF911 | K-12 (EK227) ΔgadW Δ(lacA-lacZ)::Cat | EF907 × EK432 |
| EF912 | K-12 (EK227) ΔgadXW Δ(lacA-lacZ)::Cat | EF908 × EK432 |
| EF919 | Δcya-1400::Km/pT18 × pT25 | EK395 × pT18 × pT25 |
| EF920 | Δcya-1400::Km/pMF497/pMF498 | This study |
| EF921 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−165 to +788) ΔgadW Δ(lacA-lacZ)::Cat | EF911 × EF615 |
| EF922 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−51 to +788) ΔgadW Δ(lacA-lacZ)::Cat | EF911 × EF663 |
| EF923 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−86 to +788) ΔgadW Δ(lacA-lacZ)::Cat | EF911 × EF666 |
| EF928 | K-12 (EK227) Δcya::Km | EK227 × EK395 |
| EF929 | K-12 (EK227) ΔgadXW::Km hns::Tn10 Δcrp::Cm | EF865 × EK437 |
| EF931 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−51 to +788) Δ(lacA-lacZ)::Cat | EF825 × EF663 |
| EF932 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−86 to +788) Δ(lacA-lacZ)::Cat | EF825 × EF666 |
| EF933 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−165 to +788) ΔgadXW Δ(lacA-lacZ)::Cat | EF912 × EF615 |
| EF934 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−51 to +788) ΔgadXW Δ(lacA-lacZ)::Cat | EF912 × EF663 |
| EF935 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−86 to +788) ΔgadXW Δ(lacA-lacZ)::Cat | EF912 × EF666 |
| EF948 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−51 to +788) ΔgadX Δ(lacA-lacZ)::Cat | EF830 × EF663 |
| EF949 | K-12 (EK227) trpDC::putPA1303-Km-gadA::lacZ(o) (−86 to +788) ΔgadX Δ(lacA-lacZ)::Cat | EF830 × EF663 |
| EF957 | K-12 (EK227) hns-206 oppC506::Tn10 | EK227 × EK437 |
| Plasmids | ||
| pT18 | pBluescript II KS containing T18 C terminus of B. pertussis cyaA | 9 |
| pT25 | pACYC184 containing T25 N terminus of B. pertussis cyaA | 9 |
| pT18-Zip | pT18 containing leucine zipper domain | 9 |
| pT25-Zip | pT25 containing leucine zipper domain | 9 |
| pCP20 | Flp recombinase | 3 |
| pMF497 | gadW-′cyaA in pT18 | This study |
| pMF498 | cyaA′-gadX in pT25 | This study |
| pMF499 | pET102/D-TOPO containing Trx-GadW-His6 | |
| pMF506 | cyaA′-gadW in pT25 | This study |
| pMF507 | yhiX-′cyaA in pT18 | This study |
| pET102/D-TOPO | His-Patch thioredoxin fusion vector | Invitrogen |