Tuning protein expression using synonymous codon libraries targeted to the 5' mRNA coding region

TY - JOUR

T1 - Tuning protein expression using synonymous codon libraries targeted to the 5' mRNA coding region

AU - Goltermann, Lise

AU - Borch Jensen, Martin

AU - Bentin, Thomas

PY - 2011/1

Y1 - 2011/1

N2 - In bacteria, the 5' mRNA coding region plays an important role in determining translation output. Here, we report synthetic sequences that when placed in the 5'-mRNA coding region, leading to recombinant proteins containing short N-terminal extensions, virtually abolish, enhance or produce intermediate expression levels of green fluorescent protein in Escherichia coli. At least in one case, no apparent effect on protein stability was observed, pointing to RNA level effects as the principal reason for the observed expression differences. Targeting a synonymous codon library to the 5' coding sequence allowed tuning of protein expression over ~300-fold with preservation of amino acid identity. This approach is simple and should be generally applicable in bacteria. The data support that features in the 5' mRNA coding region near the AUG start codon are key in determining translation output and hence is important to recombinant and, most certainly, endogenous gene expression.

AB - In bacteria, the 5' mRNA coding region plays an important role in determining translation output. Here, we report synthetic sequences that when placed in the 5'-mRNA coding region, leading to recombinant proteins containing short N-terminal extensions, virtually abolish, enhance or produce intermediate expression levels of green fluorescent protein in Escherichia coli. At least in one case, no apparent effect on protein stability was observed, pointing to RNA level effects as the principal reason for the observed expression differences. Targeting a synonymous codon library to the 5' coding sequence allowed tuning of protein expression over ~300-fold with preservation of amino acid identity. This approach is simple and should be generally applicable in bacteria. The data support that features in the 5' mRNA coding region near the AUG start codon are key in determining translation output and hence is important to recombinant and, most certainly, endogenous gene expression.

U2 - 10.1093/protein/gzq086

DO - 10.1093/protein/gzq086

M3 - Journal article

C2 - 21047874

SN - 1741-0126

VL - 24

SP - 125

EP - 129

JO - Protein Engineering, Design and Selection

JF - Protein Engineering, Design and Selection

IS - 1-2

ER -