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A cell‐free protein synthesis system as an investigational tool for the translation stop processes
Author(s) -
Kang Taek Jin,
Woo Ji Hyoung,
Song Hui Kyoung,
Ahn Jin Ho,
Kum Jae Wook,
Han Jin,
Choi Cha Yong,
Joo Hyun
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)02625-x
Subject(s) - translation (biology) , protein biosynthesis , cell free system , cell free protein synthesis , transfer rna , translation system , cysteine , escherichia coli , amino acid , eukaryotic translation , stop codon , biochemistry , cell , computational biology , biology , chemistry , microbiology and biotechnology , rna , messenger rna , gene , enzyme , in vitro
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.