Assessing translational efficiency by a reporter protein co-expressed in a cell-free synthesis system

Anal Biochem. 2017 Feb 1:518:139-142. doi: 10.1016/j.ab.2016.11.019. Epub 2016 Nov 29.

Abstract

We demonstrate the use of a cell-free protein synthesis system as a convenient tool for assessing the relative translational efficiencies of genes. When sfGFP was used as a common reporter gene and co-expressed with a series of target genes, the intensities of sfGFP fluorescence from the co-expression reactions were highly correlated with the individual expression levels of the co-expressed genes. The relative translational efficiencies of genes estimated by this method were reproducible when the same genes were expressed in transformed Escherichia coli, suggesting that this method could be used as a universal tool for prognostic assessment of translational efficiency.

Keywords: Assessment of gene expression; Cell-free protein synthesis; Co-expression; Translational efficiency.

MeSH terms

  • Cell-Free System / chemistry
  • Cell-Free System / metabolism
  • Escherichia coli / chemistry*
  • Escherichia coli / metabolism
  • Genes, Reporter*
  • Green Fluorescent Proteins / biosynthesis*
  • Green Fluorescent Proteins / chemistry
  • Protein Biosynthesis*

Substances

  • Green Fluorescent Proteins