High-Throughput Engineering of Initial Coding Regions for Maximized Production of Recombinant Proteins
2017
Park, Y.J., Chungnam National University, Daejeon, Republic of Korea | Lee, K.H., Chungnam National University, Daejeon, Republic of Korea | Baek, M.S., Chungnam National University, Daejeon, Republic of Korea | Kim, D.M., Chungnam National University, Daejeon, Republic of Korea
We present a method to rapidly determine the optimal initial sequences for recombinant protein production. This method relies on the competitive translation of two genes in a reaction mixture for cell-free protein synthesis. Genes from a library with randomized +2/+3 codons were individually co-expressed with superfolder green fluorescent protein (sfGFP) in this reaction mixture. As a result of competitive translation, the intensity of sfGFP fluorescence was inversely proportional to the relative translational efficiency of the co-expressed gene. Using this simple method, we identified the +2/+3 codons that markedly enhanced production of recombinant human erythropoietin compared with its wild-type codons. These +2/+3 codons also effectively enhanced protein production in Escherichia coli, indicating that this method is a useful tool to design optimal gene constructs for industrial production of recombinant proteins.
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