Fig. 2: Preparation of the hybrid-SL tRNA set.
a Exclusion of tRNASer and tRNALeu from the E. coli natural tRNA extract. The length of E. coli natural tRNAs is shown in the graph on the left. White bars represent tRNAs with 85 or more nucleotides, specifically tRNASer, tRNALeu, and tRNATyr. In contrast, gray bars represent other tRNAs with 77 or less nucleotides. Through denaturing PAGE, these two tRNA groups can be distinguished into two separate bands: band I and band II. tRNAs found in band II were subsequently extracted from the gel and are referred to as the ΔSLY natural tRNA set. The right panel shows the denaturing PAGE analysis of the natural tRNA extract, ΔSLY natural tRNA set, and IVT tRNAs (n = 1). Lanes 1 and 2 represent the tRNA extract before and after gel purification, labeled as intact and ΔSLY natural tRNA set, respectively. Lanes 3–6 represent IVT tRNASer, tRNALeu, tRNATyr, and tRNAPhe. The gel image was cropped only for this figure. b Construction of the hybrid-SL tRNA set. The ΔSLY natural tRNA set and IVT tRNAs, including chimeric tRNASerGAG, chimeric tRNALeuGGA, and tRNATyr were combined to prepare the hybrid-SL tRNA set. Abbreviations: IVT in vitro transcribed, E. coli Escherichia coli. Source data are provided as a Source Data file.
