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. 2020 Jan 10;48(4):e22. doi: 10.1093/nar/gkz1204

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Scheme 1. — Reassembling GFP by in vitro trans-translation. 1: Canonical translation of sfGFP1-10 mRNA lacking a stop codon (red cross). 2: Due to the absence of termination signal, ribosome is stalled. Two tRNAs are in the P (yellow) and E (purple) sites, and the A site is empty. 3: The tmRNAGFP11-SmpB complex binds to the stalled ribosome, and SmpB’s C-terminal tail recognizes the empty A site. 4: The translation restarts thanks to the tmRNAGFP11 MLD, which encodes the missing eleventh domain of the sfGFP (green tmRNA section) and therefore adds this beta-strand to the incomplete sfGFP1-10. 5: The process ends when the tmRNA stop codon (red star) is reached. The complete sfGFP is released and becomes fluorescent. The 50S and 30S subunits are dissociated to be reused, and the tmRNAGFP11–SmpB complex is recycled.