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. Author manuscript; available in PMC: 2021 Apr 28.
Published in final edited form as: Nature. 2020 Oct 28;587(7835):663–667. doi: 10.1038/s41586-020-2854-z

Figure 8. Extended Data Figure 8.

Figure 8

Plasmid-encoded trans-active ribozyme. a. Schematic depiction of F30-Broccoli-trans (FBT)-MTR1 with specific binding arms for E.coli tRNA-Asp (A58). b. Primer extension stop assays confirm the activity and specificity of the FBT-MTR1 in vitro transcript. Left: full gel image shown in Fig. 4b for FBT-MTR1 reacted with m6G and BG on total E.coli tRNA. Right: primer extension on BG-treated sample with six different E.coli tRNA-specific primers (P4-P9), repeated twice. c. LC-MS analysis after digestion of total E.coli tRNA treated with FBT-MTR1 and BG (same sample as for right gel image). EIC for MH+ 282.11 (methylated adenosines) and 298.11 (methylated guanosines) demonstrate the presence of natural tRNA modifications. EIC 358.11 in comparison to reference nucleosides bn1A and bn6A shows bn-modified adenosines produced by FBT-MTR1.