Development of a rapid and productive cell-free protein synthesis system |
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Authors: | Dong-Myung Kim Cha-Yong Choi Jin-Ho Ahn Tae-Wan Kim Nam-Young Kim In-Suk Oh Chang-Gil Park |
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Institution: | 1. Department of Fine Chemical Engineering and Chemistry, Chungnam National University, 305-764, Daejeon, Korea 2. School of Chemical and Biological Engineering, College of Engineering, Seoul National University, 151-742, Seoul, Korea 3. Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University, 151-742, Seoul, Korea
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Abstract: | Due to recent advances in genome sequencing, there has been a dramatic increase in the quantity of genetic information, which
has lead to an even greater demand for a faster, more parallel expression system. Therefore, interest in cell-free protein
synthesis, as an alternative method for high-throughput gene expression, has been revived. In contrast toin vivo gene expression methods, cell-free protein synthesis provides a completely open system for direct access to the reaction
conditions. We have developed an efficient cell-free protein synthesis system by optimizing the energy source and S30 extract.
Under the optimized conditions, approximately 650 μg/mL of protein was produced after 2 h of incubation, with the developed
system further modified for the efficient expression of PCR-amplified DNA. When the concentrations of DNA, magnesium, and
amino acids were optimized for the production of PCR-based cell-free protein synthesis, the protein yield was comparable to
that from the plasmid template. |
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