Fig. 6: Detailed characterization of pCNF-RS variants.
From: Integrating protein language models and automatic biofoundry for enhanced protein evolution
a Yield of sfGFP incorporated with pAcF aided by pCNF-RS and its variants. +, cell culture in presence of pAcF; -, cell culture in absence of pAcF. The concentration of purified sfGFP in presence of pAcF was 0.1 mg/mL, 0.6 mg/mL and 1.7 mg/mL for pCNF-RS, M-R3 and M-R4, respectively. b Molecular weight determination of the protein sfGFP-pAcF. Expected molecular weight is 27,832.0 Da and the peak of mass 27,831.0 Da was observed. c Efficiency of pCNF-RS and its variants suppressing multiple amber codons. Data are presented as mean values ± SD over three independent repetitions. d Incorporation efficiency of various ncAAs by pCNF-RS, M-R3 and M-R4. Data are presented as mean values ± SD over three independent repetitions. Structures of ncAAs tested are shown. e Expression of transketolase and nanobody with ncAAs incorporated by pCNF-RS and M-R4. The protein concentration was 1.8 mg/mL and 12 mg/mL for TK Y385pCNF expressed with pCNF-RS and M-R4 in presence of pCNF, respectively. Three independent experiments were conducted to confirm the results of Fig. 6a, e. Source data are provided as a Source Data file.
