Fig. 3.
Gene coupling in circuits utilising different ribosome pools. Simulations of the steady-state concentrations of RFP and GFP normalised by the maximum protein production achieved across the o-rRNA transcription rates tested. ωGFP = 100 and ωRFP = 1 to 103 mRNAs per min. o-rRNA production (ωρ) was simulated at the rates shown. Experimental data was produced by inducing RFP using AHL from 0 to 20 nM. Points are mean steady-state fluorescence ± 1 SD normalised by maximum GFP expression obtained across different levels of IPTG treatment. N = 3. Raw data is shown in Supplementary Fig. 8. The isocost line is fit to the mean fluorescence as determined by FACS from cultures during mid-exponential growth (between 3 and 5 h post-induction dependent on the strain and circuit) and gradients calculated shown in Supplementary Fig. 9. o-ribosome production was induced using three different IPTG concentrations 0.1, 0.2 and 0.5 mM as shown. a Allocation of ribosomes in b, c. b Simulations of the circuit using host ribosomes for RFP expression and o-ribosome for GFP expression. c In vivo coupling of the h-RFP-o-GFP circuit. FACS profiles are shown in Supplementary Fig. 14. d Allocation of ribosomes in e, f. e Simulations of the circuit using the o-ribosome for RFP expression and host pool for GFP. f In vivo coupling of the o-RFP-h-GFP circuit. The inset shows the data on an expanded x-axis. FACS profiles are shown in Supplementary Fig. 15