EC100D pir+ and pir-116 E. coli strain, competent cells

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Rescue cloning of EZ-Tn5 transposed elements

• Rescue transposition mutants created using the EZ-Tn5 R6K?oriKan-2> Insertion Kit
  or Tnp Transposome? Kit

• Propagate plasmids tagged with the EZ-Tn5? R6K?oriKan-2> Insertion Kit

The TransforMax EC100D pir+ Electrocompetent E. coli and TransforMax EC100D pir-116 ElectrocompetentE. coli each constitutively express the π protein (the pir gene product) for replication of plasmids containing the R6Kγ origin of replication (R6Kγori). The cells are derived from Epicentre's TransforMax? EC100? Electrocompetent E. coliby P1 phage transduction with a strain containing the pir+ or pir-116 gene linked to a dihydrofolate reductase (DHFR) marker.

The TransforMax EC100D pir+ cells will maintain R6Kγori containing plasmids at approximately 15 copies per cell,1 for cloning of potentially toxic or unstable DNA sequences. The TransforMax EC100D pir-116 cells are for high-copy propagation of up to 250 rescue plasmid copies per cell.1

Benefits

• Accepts large clones for unbiased propagation and stability of large rescue clones.

• lacZΔM15 for blue/white screening of recombinants.

• Restriction minus [mcrA, Δ(mrr-hsdRMS-mcrBC)] enables efficient cloning of methylated DNA.

• Endonuclease minus (endA1) to ensure high yields of DNA.

• Recombination minus (recA1) for greater stability of large cloned inserts.

Genotypes

TransforMax EC100D pir+ Electrocompetent E. coli:
F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) nupG pir+(DHFR)

TransforMax EC100D pir-116 Electrocompetent E. coli:
F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) nupG pir-116(DHFR)

Transformation Efficiency
Greater than 5 × 109 colony forming units (cfu)/μg with pR6Kan control DNA.

References

1. Metcalf, W.W. et al. (1994) Gene138, 1.