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. 2018 Mar 26;8:5166. doi: 10.1038/s41598-018-23201-z

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Misacylation of orthogonal tRNA for genetic code expansion. (a) Target protein cRNA containing a site-directed stop codon (nonsense codon) is subsequently suppressed by an orthogonal tRNA - THG73 or pyrrolysine tRNA (pylT). The grey highlighted nucleotides “CA” represent ligated pdCpA (see methods). (b) Co-injection of tRNA and cRNA into the Xenopus oocyte enables electrophysiological two-electrode voltage clamp characterization of ion channels and receptors containing ncAA.