title: Enhancing the efficiency of cell-free protein synthesis through the polymerase-chain-reaction-based addition of a translation enhancer sequence and the in situ removal of the extra amino acid residues.
abstract: A method for the rapid generation of intact proteins in a cell-free protein synthesis system was developed. The productivity of the recombinant proteins from the polymerase-chain-reaction-amplified templates was enhanced remarkably using an optimized translation enhancer sequence. The extra amino acid residues derived from the translation enhancer sequence were effectively removed by utilizing the appropriate detergent and peptide cleavage enzyme in the reaction mixture. These results demonstrate the versatility of cell-free protein synthesis in providing optimized and customized reaction conditions for the efficient production of the desired proteins.
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Jeong-Mi
authors.initials: JM
authors.lastname: Son
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Ahn
authors.forename: Mi-Yeon
authors.initials: MY
authors.lastname: Hwang
authors.forename: Chang-Gil
authors.initials: CG
authors.lastname: Park
authors.forename: Cha-Yong
authors.initials: CY
authors.lastname: Choi
authors.forename: Dong-Myung
authors.initials: DM
authors.lastname: Kim
mesh_terms: D002474:Cell-Free System; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D018387:Codon, Initiator / Q000737:chemistry*; D017931:DNA Primers; D015951:Factor Xa / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis; D011994:Recombinant Proteins / Q000138:chemical synthesis*
chemical_list: D018387:Codon, Initiator; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D015500:Chloramphenicol O-Acetyltransferase; D015951:Factor Xa