High-level periplasmic expression in Escherichia coli using a eukaryotic signal peptide: importance of codon usage at the 5' end of the coding sequence.
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.
Humphreys
D P
DP
Celltech-Chiroscience, 216 Bath Road, Slough, Berkshire, SL1 4EN, United Kingdom. dhumphre@celltech.co.uk
Sehdev
M
M
Chapman
A P
AP
Ganesh
R
R
Smith
B J
BJ
King
L M
LM
Glover
D J
DJ
Reeks
D G
DG
Stephens
P E
PE
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
D003062:Codon; D007140:Immunoglobulin Fab Fragments; D021382:Protein Sorting Signals; D012333:RNA, Messenger