Fig. 2.

CRISPR Cas9 mediated integration of KIV pathway into E. coli genome (A) General outline of integration of KIV pathway and curing of plasmids (B) Screening of transformed colonies by PCR to verify upstream junction (5′) and (C) downstream junction 3’. L represents 10 kb ladder; expected sizes for correct integration are represented by arrows. PCR products are separated on 0.8% agarose gel. ‘C’ represents control (without integration) Pathway is showing 80% integration efficiency (D) PCR amplification using primers (MB67/MB68) flanking outside of the integrated region to confirm whole pathway integration 5′-3’ (lane 3).