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. 2019 Aug 19;9:11971. doi: 10.1038/s41598-019-48357-0

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© The Author(s) 2019

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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Identification of PylRS variants for the incorporation of ncAA 1. (A) (Left plot) Fluorescence intensity and forward scatter of E. coli cells harboring pEVOL303_Lib in the absence of ncAAs analyzed by FACS. (Right plot) Cells harboring pEVOL303_Lib were grown in the presence of 1. Fluorescent cells (red) within the sort gate (red line) were separated from nonfluorescent cells. (B) Survival rate of cells after FACS in LB medium (1) and conditioned medium (2). Results were obtained from five independent experiments; standard deviation is indicated by error bars. (C) Fluorescence intensities of mTFP1_TAG128 expressed with 1 or without – in the present of seven different PylRS variants (mutations are shown).