In vitro suppression of two different stop codons

Eden Ozer, Yonatan Chemla, Orr Schlesinger, Haim Yuval Aviram, Inbal Riven, Gilad Haran, Lital Alfonta

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)1065-1073
Number of pages9
JournalBiotechnology and Bioengineering
Volume114
Issue number5
DOIs
StatePublished - 1 May 2017

Keywords

  • Förster resonance energy transfer
  • cell-free protein synthesis
  • dual genetic code expansion
  • orthogonal translation systems

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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