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Table 2 Strains and plasmids used in this study.

From: Generation of a restriction minus enteropathogenic Escherichia coli E2348/69 strain that is efficiently transformed with large, low copy plasmids

Strain/Plasmid Description Source/Reference
Strains   
E2348/69 Wild-type EPEC strain
EPEC (O127:H6) isolated from an infant with gastroenteritis
[27]
MC4100 Wild-type E. coli K-12 lab strain
F- araD139 Δ(argF-lac)U169 rpsL150(StrR) relA1 flbB5301 deoC1 ptsF25 rbsR
[28]
TOP10 cells Commercially available competent cells for cloning
F- mcrA Δ(mrr-hsdRMS-mcfBC) φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu) 7697 galU galK rpsL endA1 nupG (StrR)
Invitrogen Canada Inc.
TAM1 λpir Commercially available competent cells of E. coli λlysogen that provide all trans acting and mobilization factors required for the replication and mobilization of λPi dependent plasmids
mcrA Δ(mrr-hsdRMS-mcfBC) φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu) 7697 galU galK rpsL endA1 nupG λpir
Active Motif
SM10 λpir E. coli SM10 λlysogen that provide all trans acting and mobilization factors required for the replication and mobilization of λPi dependent plasmids [29]
NH1 TOP10 (pUC19 ΔhsdR) This study
NH3 SM10 λpir (pCVD442 ΔhsdR) This study
NH4 E2348/69 ΔhsdR This study
LP69 MC4100 nadA::Tn10 Lab stock
Plasmids   
pACYC184 5 kb low copy number cloning vector [16]
pUC19 Cloning vector Invitrogen Canada Inc.
pUC19 ΔhsdR pUC19 carrying ΔhsdR construct This study
pCVD442 Cloning vector requiring λ Pi protein to replicate, carries sacB for negative selection (AmpR) [10]
pCVD442 ΔhsdR pCVD442 carrying ΔhsdR construct This study
pJW15 Broad host range promoterless lux reporter plasmid [3, 13]
pLAFR1 Low copy number, broad host range plasmid [17]
pNLP10 Low copy number lux reporter plasmid, pSC101 origin [3, 13]
  1. a. Transformation efficiencies are expressed as the (number of transformants/number of viable cells)/ug DNA. All experiments were repeated at least three times, and one representative experiment is shown. b. Transformation efficiencies observed with plasmid isolated from E. coli K-12. c. Transformation efficiencies observed with plasmid isolated from E2348/69ΔhsdR.