EC100D pir+ ，EC100D pir-116大肠杆菌菌株感受态细胞-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

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

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

Rescue cloning of EZ-Tn5 transposed elements

• Rescue transposition mutants created using the EZ-Tn5 R6KɣoriKan-2> Insertion Kit
  or Tnp Transposome™ Kit

• Propagate plasmids tagged with the EZ-Tn5™ R6KɣoriKan-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.