Fig. 2. Supplemented translation machinery partially enhances protein expression.
a Graphical representation of the supplementation of purified translation factors to CFPS reactions assembled using BL-EWCE and BL-CFPWCE. Two different mixtures of purified translation machinery were produced, one with 11 IET (initiation, elongation, termination factors) and 1 AAT (aminoacyl-tRNA transferases) and the other with 11 IET and 23 AAT proteins. b Expression yield of BL-EWCE increases proportionally with the supplementation of purified translation machinery. The addition of 10 and 13 μg of purified translation factors to a 10 μL CFPS reaction increases protein yield by ~1.5-fold. Supplementation of up to 18 μg of purified translation factors increases protein yield by approximately twofold. Reaction assembly, purified protein supplementation, and incubation conditions are described in “Methods, Sections M3 and M6”. Data are presented as mean values and error bars represent s.d. (n = 4 independent experiments). Standard two-tail t-test. c Expression yield of BL-CFPWCE increases proportionally with the supplementation of purified translation machinery. The addition of 8 and 12 μg of purified translation factors to a 10 μL CFPS reaction increases protein yield by approximately twofold. Supplementation of up to 17 μg of purified protein does not result in any further increase. Reaction assembly, purified protein supplementation, and incubation conditions are described in “Methods, Sections M3 and M6”. Data are presented as mean values and error bars represent s.d. (n = 4 independent experiments). Standard two-tail t-test. Source data for b, c are provided as a Source Data file.