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Towards Reassigning the Rare AGG Codon in Escherichia coli
Author(s) -
Zeng Yu,
Wang Wei,
Liu Wenshe R.
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201400075
Subject(s) - escherichia coli , genetic code , transfer rna , codon usage bias , lysine , stop codon , mutagenesis , amino acid , biology , computational biology , genetics , biochemistry , chemistry , mutation , gene , rna , genome
The rare AGG codon in Escherichia coli has been reassigned to code non‐canonical amino acids (ncAAs) by using the PylRS‐ ${{\rm tRNA}{{{\rm Pyl}\hfill \atop {\rm CCU}\hfill}}}$ pair. When N ε ‐ alloc‐lysine was used as a PylRS substrate, almost quantitative occupancy of N ε ‐ alloc‐lysine at an AGG codon site was achieved in minimal medium. ncAAs can be potentially incorporated at the AGG codon with varying efficiencies, depending on their activities towards corresponding enzymes. As AGG is a sense codon, the approach reported here resolves the typical low ncAA incorporation issue that has been associated with ncAA mutagenesis and therefore allows bulk preparation of proteins with site‐selectively incorporated ncAAs for applications such as therapeutic protein production.