Fig. 1. A holistic synthetic biology approach to enhance cell-free systems.
a Graphical representation of the production of BL-7SWCE showing the overexpression of 11 translation factors and their influence in different stages of translation. b BL-7SWCE exhibits a higher yield than conventional cell lysates in batch CFPS reactions. We expressed deGFP encoded in the plasmid pIVEX-Eps-deGFP (10 μg μL−1) using BL-7SWCE, BL-EWCE, BL-PWCE, and the commercial S30 System. Reaction assembly, incubation conditions, and deGFP quantification are described in “Methods, Sections M3 and M4”. Reactions assembled using BL-7SWCE exhibit deGFP expression approximately two- to threefold higher than the controls. Data are presented as mean values and error bars represent s.d. (n = 3 independent experiments). Standard two-tail t-test. c BL-7SWCE exhibit approximately twofold more deGFP expression than the controls in semicontinuous exchange reactions. Assembly of the reactions and deGFP quantification are described in “Methods, Sections M4 and M5”. Data are presented as mean values and error bars represent s.d. (n = 3 independent experiments). Standard two-tail t-test. d Time series showing the expression dynamics of BL-7SWCE compared with controls under semicontinuous agitation. Reaction assembly, incubation conditions, and deGFP quantification are described in “Methods, Sections M3 and M4”. Data are presented as mean values and error bars represent 95% confidence interval (n = 4 independent experiments). Source data for b–d are provided as a Source Data file.