Cell-Free Protein Expression by a Reconstituted Transcription-Translation System Energized by Sugar Catabolism.

ABSTRACT: Cooperation between catabolism and anabolism is crucial for maintaining homeostasis in living cells. The most fundamental systems for catabolism and anabolism are the glycolysis of sugars and the transcription-translation (TX-TL) of DNA, respectively. Despite their importance in living cells, the in vitro reconstitution of their cooperation through purified factors has not been achieved, which hinders the elucidation of the design principle in living cells. Here, we reconstituted glycolysis using sugars and integrated it with the PURE system, a commercial in vitro TX-TL kit composed of purified factors. By optimizing key parameters, such as glucokinase and initial phosphate concentrations, we determined suitable conditions for their cooperation. The optimized system showed protein synthesi

SUBMITTER: Sato G 

PROVIDER: S-EPMC11243612 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature