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In vitro suppression of two different stop codons
Author(s) -
Ozer Eden,
Chemla Yonatan,
Schlesinger Orr,
Aviram Haim Yuval,
Riven Inbal,
Haran Gilad,
Alfonta Lital
Publication year - 2017
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26226
Subject(s) - genetic code , amino acid , protein biosynthesis , translation (biology) , stop codon , synthetic biology , ribosome , biology , computational biology , cell free protein synthesis , biochemistry , chemistry , gene , messenger rna , rna
Proteins play a crucial role in all living organisms, with the 20 natural amino acids as their building blocks. Unnatural amino acids are synthetic derivatives of these natural building blocks. These amino acids have unique chemical or physical properties as a result of their specific side chain residues. Their incorporation into proteins through ribosomal translation in response to one of the stop codons has opened a new way to manipulate and study proteins by enabling new functionalities, thus expending the genetic code. Different unnatural amino acids have different functionalities, hence, the ability to incorporate two different unnatural amino acids, in response to two different stop codons into one protein is a useful tool in protein manipulation. This ability has been achieved previously only in in vivo translational systems, however, with limited functionality. Herein, we report the incorporation of two different unnatural amino acids in response to two different stop codons into one protein, utilizing a cell‐free protein synthesis system. Biotechnol. Bioeng. 2017;114: 1065–1073. © 2016 Wiley Periodicals, Inc.