title: High-level periplasmic expression in Escherichia coli using a eukaryotic signal peptide: importance of codon usage at the 5' end of the coding sequence.
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.
authors.affiliation: Celltech-Chiroscience, 216 Bath Road, Slough, Berkshire, SL1 4EN, United Kingdom. dhumphre@celltech.co.uk
authors.forename: D P
authors.initials: DP
authors.lastname: Humphreys
authors.forename: M
authors.initials: M
authors.lastname: Sehdev
authors.forename: A P
authors.initials: AP
authors.lastname: Chapman
authors.forename: R
authors.initials: R
authors.lastname: Ganesh
authors.forename: B J
authors.initials: BJ
authors.lastname: Smith
authors.forename: L M
authors.initials: LM
authors.lastname: King
authors.forename: D J
authors.initials: DJ
authors.lastname: Glover
authors.forename: D G
authors.initials: DG
authors.lastname: Reeks
authors.forename: P E
authors.initials: PE
authors.lastname: Stephens
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics*; D016208:Databases, Factual; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics; D005815:Genetic Code; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009690:Nucleic Acid Conformation; D019897:Periplasm / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics*; D021381:Protein Transport; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein; D015003:Yeasts / Q000235:genetics
chemical_list: D003062:Codon; D007140:Immunoglobulin Fab Fragments; D021382:Protein Sorting Signals; D012333:RNA, Messenger