A cell‐free protein synthesis system as an investigational tool for the translation stop processes

FEBS Letters

Joo, Hyun³  Song, Hui Kyoung²  Kang, Taek Jin¹  Kum, Jae Wook²  Choi, Cha Yong¹  Han, Jin⁴  Ahn, Jin Ho²  Woo, Ji Hyoung² 

[1]School of Chemical Engineering, College of Engineering, Seoul National University, Seoul 151-742, South Korea

[2]Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University, Seoul 151-742, South Korea

[3]Department of Molecular Science and Technology/Life Sciences, Ajou University, Suwon 442-749, South Korea

[4]Department of Physiology and Biophysics, College of Medicine, Inje University, Busan 614-735, South Korea

关键词: Translation stop;    Cell-free protein synthesis;    Suppression;    Erythropoietin;    tRNA;    Unnatural amino acid;    Stop codon;    Mutagenesis;   

DOI  :  10.1016/S0014-5793(02)02625-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown