Relaxing the substrate specificity of an aminoacyl‐tRNA synthetase allows in vitro and in vivo synthesis of proteins containing unnatural amino acids

FEBS Letters

Hennecke, Hauke¹  Ibba, Michael¹ 

[1] Mikrobiologisches Institut, Eidgenössische Technische Hochschule, ETH Zentrum, Schmelzbergstrasse 7, CH-8092 Zürich, Switzerland

关键词: p-Cl-Phenylalanine;    Phenylalanine;    Phenylalanyl-tRNA synthetase;    Chaperone;    Escherichia coli;   

DOI  :  10.1016/0014-5793(95)00408-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

It has previously been demonstrated that the unnatural amino acid p-Cl-phenylalanine can be attached to tRNA^Phe by a modified phenylalanyl-tRNA synthetase with relaxed amino acid substrate specificity. We show that this modification to the translational machinery of Escherichia coli is the only requirement for the incorporation of either p-Cl- or p-Br-phenylalanine into full-length luciferase in vitro. The incorporation of p-Cl-phenylalanine was also demonstrated in vivo using a suitably modified host strain. These results represent the first description of the incorporation into a protein in vivo of an unnatural amino acid which is normally rejected by the cellular translational machinery.

【 授权许可】

Unknown