High-Throughput, Combinatorial Engineering of Initial Codons for Tunable Expression of Recombinant Proteins

@article{citeulike:2628632, abstract = {Abstract: We describe a high-throughput strategy for tuning the expression of recombinant proteins through engineering their early nucleotide sequences. After randomizing the +2 and +3 codons of the target genes, each of the variant genes was isolated in vivo and subsequently expressed using in vitro protein synthesis techniques. When several hundreds of clones were examined in parallel, it was found that expression levels of target genes varied as much as 70-fold depending on the identity of the codons in the randomized region. This broad and continuous distribution of expression levels enabled the selection of specific codon arrangements for the expression of target genes at a desired level. Furthermore, codon-dependent variations in protein expression were reproduced when the same genes were expressed in vivo. Thus, we expect that the methodology reported here could be utilized as a versatile platform for rapid expression of protein molecules at modulated levels either in vitro or in vivo.}, address = {Institute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Korea, School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea, and Department of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea}, author = {Ahn, Jin-Ho and Keum, Jung-Won and Kim, Dong-Myung }, citeulike-article-id = {2628632}, doi = {10.1021/pr700856s}, journal = {J. Proteome Res.}, keywords = {codon, expression, protein, recombinant, usage}, month = {April}, posted-at = {2008-04-04 07:14:07}, priority = {2}, title = {High-Throughput, Combinatorial Engineering of Initial Codons for Tunable Expression of Recombinant Proteins}, url = {http://dx.doi.org/10.1021/pr700856s}, year = {2008} }

TY - JOUR ID - citeulike:2628632 L3 - citeulike-article-id:2628632 TI - High-Throughput, Combinatorial Engineering of Initial Codons for Tunable Expression of Recombinant Proteins JF - J. Proteome Res. AD - Institute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Korea, School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea, and Department of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea N2 - Abstract: We describe a high-throughput strategy for tuning the expression of recombinant proteins through engineering their early nucleotide sequences. After randomizing the +2 and +3 codons of the target genes, each of the variant genes was isolated in vivo and subsequently expressed using in vitro protein synthesis techniques. When several hundreds of clones were examined in parallel, it was found that expression levels of target genes varied as much as 70-fold depending on the identity of the codons in the randomized region. This broad and continuous distribution of expression levels enabled the selection of specific codon arrangements for the expression of target genes at a desired level. Furthermore, codon-dependent variations in protein expression were reproduced when the same genes were expressed in vivo. Thus, we expect that the methodology reported here could be utilized as a versatile platform for rapid expression of protein molecules at modulated levels either in vitro or in vivo. KW - codon KW - expression KW - protein KW - recombinant KW - usage AU - Ahn, Jin-Ho AU - Keum, Jung-Won AU - Kim, Dong-Myung PY - 2008/04/04/ UR - http://dx.doi.org/10.1021/pr700856s ER -