High-level periplasmic expression in Escherichia coli using a eukaryotic signal peptide: importance of codon usage at the 5' end of the coding sequence

Protein Expr Purif. 2000 Nov;20(2):252-64. doi: 10.1006/prep.2000.1286.

Abstract

We investigated the ability of signal peptides of eukaryotic origin (human, mouse, and yeast) to efficiently direct model proteins to the Escherichia coli periplasm. These were compared against a well-characterized prokaryotic signal peptide-OmpA. Surprisingly, eukaryotic signal peptides can work very efficiently in E. coli, but require optimization of codon usage by codon-based mutagenesis of the signal peptide coding region. Analysis of the 5' of periplasmic and cytoplasmic E. coli genes shows some codon usage differences.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Codon / genetics*
  • Databases, Factual
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Gene Expression Regulation, Bacterial*
  • Genes, Bacterial / genetics
  • Genetic Code
  • Humans
  • Immunoglobulin Fab Fragments / genetics
  • Mice
  • Molecular Sequence Data
  • Mutagenesis / genetics
  • Nucleic Acid Conformation
  • Periplasm / metabolism*
  • Plasmids / genetics
  • Protein Sorting Signals / genetics*
  • Protein Transport
  • RNA Stability
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Analysis, Protein
  • Yeasts / genetics

Substances

  • Codon
  • Immunoglobulin Fab Fragments
  • Protein Sorting Signals
  • RNA, Messenger