Overcoming codon bias: a method for high-level overexpression of Plasmodium and other AT-rich parasite genes in Escherichia coli.
Parasite genes often use codons which are rarely used in the highly expressed genes of Escherichia coli, possibly resulting in translational stalling and lower yields of recombinant protein. We have constructed the "RIG" plasmid to overcome the potential codon-bias problem seen in Plasmodium genes. RIG contains the genes that encode three tRNAs (Arg, Ile, Gly), which recognise rare codons found in parasite genes. When co-transformed into E. coli along with expression plasmids containing parasite genes, RIG can greatly increase levels of overexpressed protein. Codon frequency analysis suggests that RIG may be applied to a variety of protozoan and helminth genes.
Baca
A M
AM
Department of Bioengineering, Howard Hughes Medical Institute, Biomolecular Structure Center, Box 357742, University of Washington, Seattle, WA 98195, USA.
Hol
W G
WG
D020856:AT Rich Sequence / Q000235:genetics*; D000818:Animals; D003062:Codon; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005786:Gene Expression Regulation; D005787:Gene Frequency; D010957:Plasmids / Q000235:genetics; D010961:Plasmodium / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014162:Transfection
D003062:Codon