Fig. 2: Protein expression with a single plasmid encoding all 21 tRNA genes. | Nature Communications

Fig. 2: Protein expression with a single plasmid encoding all 21 tRNA genes.

From: Continuous in situ synthesis of a complete set of tRNAs sustains steady-state translation in a recombinant cell-free system

Fig. 2

a A schematic describing the preparation of mature tRNAs from a circular template nicked with Nt.BspQI. Created in BioRender. Maerkl, S. (2025) https://BioRender.com/eh7ck4h. b UREA-PAGE analysis of tRNAs transcribed from nicked and un-nicked plasmid. E. coli tRNAs are also shown for comparison. c Schematic describing the approach to test the activity of IVT tRNAs transcribed from a nicked plasmid in ΔtRNA PURE system (n = 4 for 0 and 0.5 μg/μl tRNAnicked, n = 5 for 0.5 and 1 μg/μl tRNAunnicked, 1 μg/μl tRNAnicked). The bar plot shows the expression of sfGFP with various concentrations of IVT tRNA added to the ΔtRNA PURE system. Created in BioRender. Maerkl, S. (2025) https://BioRender.com/eh7ck4h. d Approach for testing the activity of nicked plasmid added directly to theΔtRNA PURE system for in situ tRNA transcription (n = 4). Created in BioRender. Maerkl, S. (2025) https://BioRender.com/eh7ck4h. The bar plot shows the expression of sfGFP with the indicated concentration of nicked plasmid added to the ΔtRNA PURE system. Each dot represents a data point from an independent replicate. Bars and error bars represent the mean and standard deviation. Source data are provided as a Source Data file.

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