Streamlined cell-free protein synthesis from sequence information |
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Authors: | Jin-Ho Ahn Kyung-Ho Lee Jae-Won Shim Eun Yeol Lee Dong-Myung Kim |
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Institution: | 1. Department of Fine Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764, Korea 2. Department of Biochemistry and Molecular Biology, College of Medicine, Hanyang University, Seoul, 133-791, Korea 3. Department of Chemical Engineering, Kyung Hee University, Yongin, 446-701, Korea
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Abstract: | In this study, we describe a cell-free protein synthesis consolidated with polymerase chain reaction (PCR)-based synthetic gene assembly that allows for streamlined translation of genetic information. In silico-designed fragments of target genes were PCR-assembled and directly expressed in a cell-free synthesis system to generate functional proteins. This method bypasses the procedures required in conventional cell-based gene expression methods, integrates gene synthesis and cell-free protein synthesis, shortens the time to protein production, and allows for facile regulation of gene expression by manipulating the oligomer sequences used for gene synthesis. The strategy proposed herein expands the flexibility and throughput of the protein synthesis process, a fundamental component in the construction of synthetic biological systems. |
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