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. 2016 Sep 26;45(4):2040–2050. doi: 10.1093/nar/gkw854

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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 and analysis of 3GC mutants of tRNA^fMet. (A) Sequences of the anticodon stem loops of the mutants used in this study. (B) Expression analysis of the mutant tRNA^fMet. Approximately 10 μg of total tRNA preparations from E. coli KL16, or KL16ΔmetY::cm (retains tRNA^fMet1, as indicated) harboring plasmids containing the mutant metY genes were separated on native polyacrylamide gel electrophoresis (PAGE), transferred onto nytran membrane and hybridized with D-loop ini tRNA probe (complementary to 2–27 nt position) common to all tRNA^fMet. tRNA^Met (detected by met elongator D loop probe) serves as internal loading control. The two conformer of mid GU and au/gu/au tRNAs are marked by asterisks (*), and the band corresponding to au/gu/gu has been indicated by an arrowhead. Longer exposure of lane 15 has also been shown.