Transformation of the single deletions strains E. coli strains KRX ∆alr and DH5alpha ∆alr with the plasmid pRedET containg the Recombinase using the Genebridge RedET-System protocol.
Transformation of the oligonucleotide containing the flanking sites for the deletion of dadX. The successful replacement of the catabolic alanine racemase was verified via kanamycin selection and
Colony PCR (dadX_Ec_control1, dadX_Ec_control2)
Annealing temperature: 55 °C
Bands as expected (3004 bp)
Resulting in the genotype DH5aplha ∆alrkan:dadX, while the dadX deletion in the KRX strain was not successful.
Transformation of the oligonucleotide containing the flanking sites for the deletion of dadX into the KRX ∆alr. The successful replacement of the katabolic alanine racemase was verified via kanamycin selection and
Colony PCR (dadX_Ec_control1, dadX_Ec_control2)
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