title: Genetic Code Expansion by Degeneracy Reprogramming of Arginyl Codons.
abstract: The genetic code in most organisms codes for 20 proteinogenic amino acids or translation stop. In order to encode more than 20 amino acids in the coding system, one of stop codons is usually reprogrammed to encode a non-proteinogenic amino acid. Although this approach works, usually only one amino acid is added to the amino acid repertoire. In this study, we incorporated non-proteinogenic amino acids into a protein by using a sense codon. As all the codons are allocated in the universal genetic code, we destroyed all the tRNA(Arg) in a cell-free protein synthesis system by using a tRNA(Arg) -specific tRNase, colicin D. Then by supplementing the system with tRNACCU , the translation system was partially restored. Through this creative destruction, reprogrammable codons were successfully created in the system to encode modified lysines along with the 20 proteinogenic amino acids.
authors.affiliation: Department of Chemical and Biochemical Engineering, Dongguk University-Seoul, Seoul, 100-715, South Korea.
authors.forename: Ki Baek
authors.initials: KB
authors.lastname: Lee
authors.affiliation: Department of Chemical and Biochemical Engineering, Dongguk University-Seoul, Seoul, 100-715, South Korea.
authors.forename: Chen Yuan
authors.initials: CY
authors.lastname: Hou
authors.affiliation: Department of Chemical and Biochemical Engineering, Dongguk University-Seoul, Seoul, 100-715, South Korea.
authors.forename: Chae-Eun
authors.initials: CE
authors.lastname: Kim
authors.affiliation: Department of Fine Chemical Engineering and Applied Chemistry, Chungnam National University, Daejun, 305-764, South Korea.
authors.forename: Dong-Myung
authors.initials: DM
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo, 113-0033, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Suga
authors.affiliation: Department of Chemical and Biochemical Engineering, Dongguk University-Seoul, Seoul, 100-715, South Korea. taekkang@dongguk.edu.
authors.forename: Taek Jin
authors.initials: TJ
authors.lastname: Kang
mesh_terms: D001120:Arginine / Q000235:genetics*; D003062:Codon; D003087:Colicins / Q000378:metabolism; D019020:Directed Molecular Evolution; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005815:Genetic Code; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D012347:RNA, Transfer, Arg / Q000235:genetics / Q000378:metabolism
chemical_list: D003062:Codon; D003087:Colicins; D012347:RNA, Transfer, Arg; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D001120:Arginine
keywords: colicin D; degenerate codons; mutagenesis; protein engineering; tRNA