Fig. 4
From: Cell-free prediction of protein expression costs for growing cells
Predicting the burden of operon designs for the beta-carotene biosynthesis pathway. a Diagram of the beta-carotene pathway and the γ-values for the four enzyme-encoding sequences as measured by the cell lysate capacity assay (see Supplementary Fig. 7). The operon is designed with partially randomised RBS sequences and one of three promoters: BBa_J23113 (weak), BBa_J23106 (medium), or BBa_J23100 (strong). b Model-predicted burden of each operon design compared to the measured in vivo capacity of E. coli expressing the operons with or without an inactivating mutation in the crtE gene (prediction method described in Supplementary Fig. 8). The orange intensity in each circle represents the measured beta-carotene level for each strain (see Supplementary Fig. 9). Error bars show standard error of three independent repeats. c Model-predicted burden of each operon design compared to the measured in vivo capacity of E. coli expressing the active pathway (same data as b, non-mutated pathway on left). The diagonal dot line represents equality between predicted and measured normalised in vivo capacity. Grey bars indicate the difference between the predicted and measured normalised in vivo capacity of the 17 operons. Right plot compares the relative differences between predicted and measured normalised capacity for the 17 operons and the strain-only control. Operons are ranked from low to high-in vivo capacity values. Values are normalised to the capacity obtained with capacity monitor plasmid alone
