Development of a Rapid and Productive Cell-free Protein Synthesis System

• Published in: Biotechnology and bioprocess engineering Vol. 11; no. 3; pp. 235 - 239
• Main Authors: Kim, Dong-Myung, Choi, Cha-Yong, Ahn, Jin-Ho, Kim, Tae-Wan, Kim, Nam-Young, Oh, In-Suk, Park, Chang-Gil
• Format: Journal Article
• Language: Korean
• Published: 한국생물공학회 01.06.2006
• Subjects: 생물공학; cell-free protein synthesis; ATP regeneration; PCR
• ISSN: 1226-8372; 1976-3816

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.