Issue 6, 2016

Performance of optimized noncanonical amino acid mutagenesis systems in the absence of release factor 1

Abstract

Site-specific incorporation of noncanonical amino acids (ncAAs) into proteins expressed in E. coli using UAG-suppression competes with termination mediated by release factor 1 (RF1). Recently, unconditional deletion of RF1 was achieved in a genomically recoded E. coli (C321), devoid of all endogenous UAG stop codons. Here we evaluate the efficiency of ncAA incorporation in this strain using optimized suppression vectors. Even though the absence of RF1 does not benefit the suppression efficiency of a single UAG codon, multi-site incorporation of a series of chemically distinct ncAAs was significantly improved.

Graphical abstract: Performance of optimized noncanonical amino acid mutagenesis systems in the absence of release factor 1

Supplementary files

Article information

Article type
Communication
Submitted
28 Jän 2016
Accepted
23 Mär 2016
First published
30 Mär 2016

Mol. BioSyst., 2016,12, 1746-1749

Author version available

Performance of optimized noncanonical amino acid mutagenesis systems in the absence of release factor 1

Y. Zheng, M. J. Lajoie, J. S. Italia, M. A. Chin, G. M. Church and A. Chatterjee, Mol. BioSyst., 2016, 12, 1746 DOI: 10.1039/C6MB00070C

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