Tuning of Recombinant Protein Expression in Escherichia coli by Manipulating Transcription, Translation Initiation Rates, and Incorporation of Noncanonical Amino Acids

Orr Schlesinger, Yonatan Chemla, Mathias Heltberg, Eden Ozer, Ryan Marshall, Vincent Noireaux, Mogens Høgh Jensen, Lital Alfonta

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Protein synthesis in cells has been thoroughly investigated and characterized over the past 60 years. However, some fundamental issues remain unresolved, including the reasons for genetic code redundancy and codon bias. In this study, we changed the kinetics of the Eschrichia coli transcription and translation processes by mutating the promoter and ribosome binding domains and by using genetic code expansion. The results expose a counterintuitive phenomenon, whereby an increase in the initiation rates of transcription and translation lead to a decrease in protein expression. This effect can be rescued by introducing slow translating codons into the beginning of the gene, by shortening gene length or by reducing initiation rates. On the basis of the results, we developed a biophysical model, which suggests that the density of co-transcriptional-translation plays a role in bacterial protein synthesis. These findings indicate how cells use codon bias to tune translation speed and protein synthesis.

Original languageEnglish (US)
Pages (from-to)1076-1085
Number of pages10
JournalACS Synthetic Biology
Volume6
Issue number6
DOIs
StatePublished - Jun 16 2017

Bibliographical note

Publisher Copyright:
© 2017 American Chemical Society.

Keywords

  • codon bias
  • genetic code expansion
  • protein translation initiation
  • rates of translation
  • transcription initiation

Fingerprint

Dive into the research topics of 'Tuning of Recombinant Protein Expression in Escherichia coli by Manipulating Transcription, Translation Initiation Rates, and Incorporation of Noncanonical Amino Acids'. Together they form a unique fingerprint.

Cite this