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. 2019 Jun 10;47(15):8207–8223. doi: 10.1093/nar/gkz503

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© The Author(s) 2019. 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 License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 10. — Ribosomal pausing at cardiovirus frameshift signals. (A) Wild type (WT) and shift site mutant (SS) TMEV and EMCV frameshift sequences were inserted into reporter plasmid pPS0. (B–H) RNAs derived from AvaII-cut plasmids were translated in WG and, after 5 min, further initiation was halted by the addition of edeine, and aliquots were removed at various times and analyzed by SDS-PAGE. Lanes M and C show markers and the expected size of the ribosomal pause product, respectively. Translations were supplemented with 1.8 μM TMEV 2A, TMEV 2A-mut, or EMCV 2A as indicated. (B) TMEV SS RNA + TMEV 2A. (C) TMEV WT RNA + TMEV 2A. (D) TMEV SS RNA + TMEV 2A-mut. (E) TMEV WT RNA + TMEV 2A-mut. (F) EMCV SS RNA + EMCV 2A. (G) EMCV SS RNA + TMEV 2A. (H) TMEV SS RNA + EMCV 2A. As well as the full-length product and the transient pausing product, a frameshift product is produced for TMEV WT RNA in the presence of TMEV 2A only.