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Table 1 Strains and plasmids used in the study

From: Role of EscU auto-cleavage in promoting type III effector translocation into host cells by enteropathogenic Escherichia coli

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

  

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