Highly Productive One-Pot System for the Incorporation of Non-Standard Amino Acids into Cell-Free Synthesized Proteins

摘要

Disclosed are methods, systems, components, and compositions related to genomically recoded and engineered organisms. The disclosed genomically recoded and engineered organisms may be used to prepare extracts for use in platforms and methods for preparing sequence defined biopolymers in vitro. In particular, the methods, systems, components, and compositions relate to genomically recoded and engineered organisms comprising a strain deficient in release factor 1 (RF-1) or a genetic homolog thereof, wherein the genomically recoded organisms have been engineered to express a heterologous RNA polymerase that may be utilized to express a target protein from a transcription template comprising a promoter for the heterologous RNA polymerase, such as bacteriophage T7 RNA polymerase.