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Table 1 Bacterial strains used in this study

From: A simple method for construction of pir+ Enterobacterial hosts for maintenance of R6K replicon plasmids

Bacterial strains

E. coli

Relevant genotype

Source or reference

DB3.1

F- gyrA462 endA1 glnV44 Δ(sr1-recA) mcrB mrr hsdS20 (rB- mB-) ara14 galK2 lacY1 proA2 rpsL20 xyl5 Δleu mtl1

Invitrogen

PIR1

F- Δ(argF-lac)169 rpoS(Am) robA1 creC510 hsdR514 endA recA1 uidA(ΔMluI)::pir-116

Invitrogen

PIR2

F- Δ(argF-lac)169 rpoS(Am) robA1 creC510 hsdR514 endA recA1 uidA(ΔMluI)::pir+

Invitrogen

RHO3

SM10 (λpir)1 Δasd::FRT ΔaphA::FRT

[19]

DH5α

F- ϕ80 lacZ ΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK- mK+) phoA glnV44

[20]

JM108

mcrA recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 Δ(lac-proAB)

[21]

MC4100

F- araD139 Δ(argF-lac)169 flhD5301 Δ(fruK-yeiR)725 (fruA25) relA1 rpsL150 rbsR22 Δ(fimB-fimE)632::IS deoC1

[22, 23]

SM10

thi-1 thr-1 leuB26 tonA21 lacY1 supE44 recA integrated RP4-2 Tcr::Mu aphA+ (Kmr)

(RP4-2 is RP4 ΔTn1)

[24]

SBr1

DH5α::mini- Tn7-pir+

This study

SBr2

JM108::mini- Tn7-pir+

This study

SBr3

MC4100::mini- Tn7-pir+

This study

SBr4

SM10::mini- Tn7-pir+

This study

MaH1

DH5α::mini-Tn7-pir-116

This study

MaH2

JM108::mini-Tn7-pir-116

This study

MaH3

MC4100::mini-Tn7-pir-116

This study

MaH4

SM10::mini-Tn7-pir-116

This study

MaH5

DH5α::mini-Tn7-pir-116- FKm

This study

RHO42

RHO3::mini-Tn7-pir-116- FKm

This study

RHO52

RHO3::mini-Tn7-pir-116-FRT

This study

S. enterica serovar Typhimurium

  

14028S

Wild Type

ATCC

SDr1

14028S::mini- Tn7-pir+

This study

SDr2

14028S::mini-Tn7-pir-116

This study

  1. 1The pir gene carried by λpir has a truncation that removes the coding region for the carboxy-terminal 30 amino acids of the π protein. Despite this truncation a λpir lysogen maintains plasmids with at the same copy number as cells carrying a wild-type pir+ gene [9].
  2. 2The pir-116 allele is dominant over the pir gene carried by the lysogenic λ phage and leads to increased copy number of oriR6Kγ containing plasmids.