Relaxing the Substrate Specificity of an Aminoacyl-tRNA Synthetase Allows in vitro and in vivo Synthesis of Proteins Containing Unnatural Amino Acids
It has previously been demonstrated that the unnatural amino acid p‐Cl‐phenylalanine can be attached to tRNAPhe 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.
Ibba, M. and Hennecke, H. (1995) Relaxing the substrate specificity of an aminoacyl-tRNA synthetase allows in vitro and in vivo synthesis of proteins containing unnatural amino acids. FEBS Lett. 364, 272-275. https://doi.org/10.1016/0014-5793(95)00408-2
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