Fig. 3: Optimization and expansion of C:G to T:A type single mutators. | Nature Communications

Fig. 3: Optimization and expansion of C:G to T:A type single mutators.

From: An orthogonal transcription mutation system generating all transition mutations for accelerated protein evolution in vivo

Fig. 3: Optimization and expansion of C:G to T:A type single mutators.

a Optimization of the PmCDA1 mutator by independently expressing UGI with strong, medium, and weak RBSs. b Analysis of on-target mutation rates of PmCDA1 mutators with three different strengths of RBSs. c Expansion of C:G to T:A type single mutators via the fusion of PmCDA1-UGI to K1F and VP4 RNAPs, respectively. To assess the on-target mutation rate, Erythromycin resistance mutant ErmC Y104S was used under the control of PK1F and PVP4 promoters, respectively. d–f Analysis of the on-target mutation rate, off-target rate, and cell viability of K1F and VP4-based C:G to T:A type single mutators. Data are presented as mean values (bars), standard errors (error bars), and individual values (black dots). n = 3, which represents three independent replicates of the experiment. For statistical analyses, two-tailed Student’s t-tests were applied. A p-value < 0.05 was considered significant. Source data are provided as a Source Data file.

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