Cell-free protein synthesis from genomically recoded bacteria enables multisite incorporation of noncanonical amino acids.

ABSTRACT: Cell-free protein synthesis has emerged as a powerful approach for expanding the range of genetically encoded chemistry into proteins. Unfortunately, efforts to site-specifically incorporate multiple non-canonical amino acids into proteins using crude extract-based cell-free systems have been limited by release factor 1 competition. Here we address this limitation by establishing a bacterial cell-free protein synthesis platform based on genomically recoded Escherichia coli lacking release factor 1. This platform was developed by exploiting multiplex genome engineering to enhance extract performance by functionally inactivating negative effectors. Our most productive cell extracts enabled synthesis of 1,780 ± 30 mg/L superfolder green fluorescent protein. Using an optimized platform, we dem

SUBMITTER: Martin RW 

PROVIDER: S-EPMC5865108 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature