Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Development of a Rapid and Productive Cell-free Protein Synthesis System

  • Kim, Dong-Myung (Department of Fine Chemical Engineering and Chemistry, Chungnam National University) ;
  • Choi, Cha-Yong (School of Chemical and Biological Engineering, College of Engineering, Seoul Natioal University) ;
  • Ahn, Jin-Ho (Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University) ;
  • Kim, Tae-Wan (School of Chemical and Biological Engineering, College of Engineering, Seoul Natioal University) ;
  • Kim, Nam-Young (School of Chemical and Biological Engineering, College of Engineering, Seoul Natioal University) ;
  • Oh, In-Suk (School of Chemical and Biological Engineering, College of Engineering, Seoul Natioal University) ;
  • Park, Chang-Gil (Department of Fine Chemical Engineering and Chemistry, Chungnam National University)
  • Published : 2006.06.30
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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 to in 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 $650{\mu}g/mL$ of protein was produced after 2h 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.

Keywords

References

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