Strains | Description | Source/comment |
---|---|---|
Wild type EPEC | EPEC strain E2348/69, streptomycin-resistant, BFP positive. | [35] |
ΔescU | escU deletion mutant | This study |
ΔsepD | sepD deletion mutant |  |
ΔsepDΔescU | Double mutant derived from ΔsepD | This study |
ΔsepD::escU(N262A) | Cis-complemented ΔsepDΔescU strain | This study |
ΔsepD::escU(P263A) | Cis-complemented ΔsepDΔescU strain | This study |
ΔsepDΔtir | Double mutant derived from ΔsepD | [35] |
ΔescNΔescU | Double mutant derived from ΔescN | This study |
SM10λpir | E. coli strain that is permissive for pRE112 replication |  |
DH5α | E. coli strain used for cloning |  |
DH5αλpir | E. coli strain used for cloning, permissive for pRE112 replication |  |
Plasmids | Â | Â |
pET21 a+ pACYC184 | T7/HIS tagged fusion vector Broad host range plasmid, P15A derived | Novagen [72] |
pETescUHIS | pET21a+ expressing EscU-HIS | This study |
pETescU(N262A)HIS | pET21a+ expressing EscU(N262A)-HIS | This study |
pETescU(P263A)HIS | pET21a+ expressing EscU(P263A)-HIS | This study |
pEscN | pACYC184 expressing EscN | [65] |
pFLAG-CTC | Cloning vector to express C-terminal FLAG fusion proteins from tac promoter | Sigma |
pJLT21 | pACYC184 expressing EscU-HIS | This study |
pJLT22 | pACYC184 expressing EscU(N262A)-HIS | This study |
pJLT23 | pACYC184 expressing EscU(P263A)-HIS | This study |
pJLT24 | pFLAG-CTC backbone vector that expresses a HA-EscU-FLAG tagged protein | This study |
pJLT25 | pFLAG-CTC backbone vector that expresses a HA-EscU(N262A)-FLAG tagged protein | This study |
pJLT26 | pFLAG-CTC backbone vector that expresses a HA-EscU(P263A)-FLAG tagged protein | This study |
pRE112 | λpir origin, suicide vector encoding sacB | [73] |
PΔescU | escU deletion fragment cloned into pRE112 suicide vector | This study |
pTir-bla | pCX341 expressing a Tir-TEM1 fusion protein | This study |