Fig. 2

Adapted from Rugbjerg et al., 2018. One “wild-type” E. coli MG1655 strain (W3110) and a reduced genome strain (MDS42) were cultivated harbouring one out of three plasmid constructs (Fig. 1A). By serially transferring samples every 10 or 5 h into fresh medium, respectively, we imitate generation numbers found in large-scale industrial bioreactors. Thereby the cultures were kept in the exponential phase throughout the experiment. Sample collection was continued until desired generation numbers (> 60) were achieved. Subsequently, all cryo-samples were re-cultivated for 72 h during which growth rate- and final L-cysteine yields were monitored. RNA extractions for comparative transcriptomics and plasmid extractions for deep plasmid sequencing were performed on the early and late generation population (EGP, LGP) samples for each strain and plasmid construct, respectively. Created with BioRender.com