title: A cell-free protein synthesis system as an investigational tool for the translation stop processes.
abstract: Using Escherichia coli cell-free protein synthesis system and aminoacylated amber suppressor tRNA, we successfully inserted an unnatural amino acid S-(2-nitrobenzyl)cysteine into human erythropoietin. Three different types of translation stop suppression were observed and each of the three types was easily discerned with SDS-PAGE. Optimal conditions were established for correct stop and programmed suppressions. Since this system differentiates proteins produced by misreading of codons from those produced by programmed suppression, we conclude that this cell-free translation system that we describe in this paper will be of a great use for future investigations on translation stop processes.
authors.affiliation: School of Chemical Engineering, College of Engineering, Seoul National University, Seoul, South Korea.
authors.forename: Taek Jin
authors.initials: TJ
authors.lastname: Kang
authors.forename: Ji Hyoung
authors.initials: JH
authors.lastname: Woo
authors.forename: Hui Kyoung
authors.initials: HK
authors.lastname: Song
authors.forename: Jin Ho
authors.initials: JH
authors.lastname: Ahn
authors.forename: Jae Wook
authors.initials: JW
authors.lastname: Kum
authors.forename: Jin
authors.initials: J
authors.lastname: Han
authors.forename: Cha Yong
authors.initials: CY
authors.lastname: Choi
authors.forename: Hyun
authors.initials: H
authors.lastname: Joo
mesh_terms: D002474:Cell-Free System / Q000378:metabolism; D018388:Codon, Terminator / Q000235:genetics* / Q000378:metabolism; D003545:Cysteine / Q000031:analogs & derivatives* / Q000378:metabolism; D004921:Erythropoietin / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016254:Mutagenesis, Insertional; D014176:Protein Biosynthesis; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*
chemical_list: D018388:Codon, Terminator; D012346:RNA, Transfer, Amino Acyl; D004921:Erythropoietin; D003545:Cysteine