Figure 5.
Scar-free single-base genome engineering. (A) Plasmids in the edited cells were cured through incubation at a high temperature, and the cas9-KmR cassette was replaced with the araBAD operon through P1 transduction. (B) The chromosomal structures in the ara operon in E. coli MG1655, HK1059, HL001, and HK1164 cells were confirmed via PCR. (C) The phenotype (Gal−Ara+) of single-base-edited HK1164 cells (=MG1655, galK T504A), as confirmed through cell culture on MacConkey agar supplemented with D-galactose or L-arabinose.
