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. 2016 May 25;44(13):6471–6481. doi: 10.1093/nar/gkw470

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© The Author(s) 2016. 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-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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Figure 1. — Generation of a RelA-stalled ribosome complex. (A) The bicistronic 2XermCL_S10K mRNA was translated in the presence of 10 μM erythromycin (ERY) in order to generate (B) ErmCL-S10K_SRC disomes. (C) ErmCL_S10K-SRC disomes were converted to monosomes by antisense DNA-mediated RNase H cleavage, as shown by sucrose density gradient centrifugation and negative stain electron microscopy (EM). (D) A-tRNA deficient ErmCL_S10K-SRCs were used as substrate for (E) RelA binding in the presence of deacylated tRNA^Lys, GDP and α, β-methylene-ATP.