The TransforMax™ EC100D™ pir Electrocompetent E. coli and TransforMax™ EC100D™ pir-116 Electrocompetent E. coli each express the p protein (the pir gene product) for replication of plasmids containing the R6Kγ origin of replication (R6Kγ ori). The cells are derived from Epicentre's® TransforMax™ E. coli Electrocompetent E. coli by P1 phage transduction with a strain containing the pir or pir-116 gene linked to a dihydrofolate reductase (DHFR) marker.
The TransforMax™ EC100D™ pir Electrocompetent E. coli 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 Electrocompetent E. coli are for high copy propagation of up to 250 rescue plasmid copies per cell.1
EZ-Tn5™ Transposons containing the R6Kγori can be introduced into genomic DNA in vivo or plasmid DNA in vitro for 'rescue' cloning applications using the EZ-Tn5™ <R6Kγori /KAN-2> Tnp Transposome or the EZ-Tn5™ <R6Kγori /KAN-2> Insertion Kit, respectively. You can also make your own R6Kγori containing EZ-Tn5™ Transposon using the EZ-Tn5™ pMOD-3 <R6Kγori /MCS> Transposon Construction Vector.
Transformation Efficiency
Greater than 1X109 colony forming units (cfu)/µg supercoiled DNA
Genotypes
TransforMax™ EC100D™ pir Electrocompetent E. coli
Maintains plasmids at approximately 250 copies per cell.
F- mcrA delta(mrr-hsdRMS-mcrBC) omega80dlacZdeltaM15 deltalacX74 recA1 endA1 araD139 delta(ara, leu)7697 galU galK γ- rpsL nupG pir-116(DHFR).
Important Phenotypes & Applications
- Expresses the <pi> Protein (pir gene product) - for rescue cloning and propagation of vectors containing the R6Kγ origin of replication
- Restriction minus - for efficient cloning of methylated genomic DNA
- Accepts large clones for unbiased propagation and stability of large rescue clones
- Endonuclease minus (endA1) to ensure high yields of plasmid clones
- Recombination minus (recA1) - for stability of large clones
- Compatible with vectors expressing the LacZ™ a-complementing peptide - for blue/white screening of recombinants
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