Figure 2

Characterization of the pKOCOMP- adk /pI- Sce I and the pCOMP- adk /pP _ara I- Sce I systems. A: Elimination of pKOCOMP-adk from adk::kan due to rhamnose-induced expression of I-Sce I nuclease from helper plasmid pI-Sce I. Red arrow: Time point for harvest and competent cell preparation. The inlet gives a better resolution of the elimination dynamics. The chromosomal adk-replacement was established using pKOCOMP-adk. B: Elimination of pCOMP-adk from adk::kan due to arabinose-induced expression of I-Sce I nuclease from helper plasmid pP _ara I-Sce I. The chromosomal adk-replacement was established using pKD46 and pCOMP-adk.