A Bacterial Strain with a Unique Quadruplet Codon Specifying Non‐native Amino Acids
- 27 May 2014
- journal article
- research article
- Published by Wiley in ChemBioChem
- Vol. 15 (12) , 1782-1786
Abstract
The addition of noncanonical amino acids to the genetic code requires unique codons not assigned to the 20 canonical amino acids. Among the 64 triplet codons, only the three nonsense “stop” codons have been used to encode non‐native amino acids. Use of quadruplet “frame‐shift” suppressor codons provides an abundant alternative but suffers from low suppression efficiency as a result of competing recognition of their first three bases by endogenous host tRNAs or release factors. Deletion of release factor 1 in a genomically recoded strain of E. coli (E. coli C321), in which all endogenous amber stop codons (UAG) are replaced with UAA, abolished UAG mediated translation termination. Here we show that a Methanocaldococcus jannaschii‐derived frame‐shift suppressor tRNA/aminoacyl‐tRNA synthetase pair enhanced UAGN suppression efficiency in this recoded bacterial strain. These results demonstrate that efficient quadruplet codons for encoding non‐native amino acids can be generated by eliminating competing triplet codon recognition at the ribosome.Keywords
This publication has 22 references indexed in Scilit:
- Evolution of multiple, mutually orthogonal prolyl-tRNA synthetase/tRNA pairs for unnatural amino acid mutagenesis in Escherichia coliProceedings of the National Academy of Sciences, 2012
- Catalyst‐Free and Site‐Specific One‐Pot Dual‐Labeling of a Protein Directed by Two Genetically Incorporated Noncanonical Amino AcidsChemBioChem, 2012
- In Vivo Incorporation of Multiple Noncanonical Amino Acids into ProteinsAngewandte Chemie International Edition, 2011
- Paralleler In‐vivo‐Einbau von mehreren nichtkanonischen Aminosäuren in ProteineAngewandte Chemie, 2011
- An Evolved Aminoacyl-tRNA Synthetase with Atypical Polysubstrate Specificity,Biochemistry, 2011
- Adding New Chemistries to the Genetic CodeAnnual Review of Biochemistry, 2010
- A Facile System for Genetic Incorporation of Two Different Noncanonical Amino Acids into One Protein in Escherichia coliAngewandte Chemie, 2010
- De Novo Generation of Mutually Orthogonal Aminoacyl-tRNA Synthetase/tRNA PairsJournal of the American Chemical Society, 2010
- Expanding the genetic code: selection of efficient suppressors of four-base codons and identification of “shifty” four-base codons with a library approach in Escherichia coli11Edited by M. GottesmanJournal of Molecular Biology, 2001
- Quadruplet codons: implications for code expansion and the specification of translation step sizeJournal of Molecular Biology, 2000