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. 2014 Dec 23;16(1):91–113. doi: 10.3390/ijms16010091

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© 2014 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

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The Ta0836 gene product is T. acidophilum Trm5. (A) 10 µg of the purified Ta0836 gene product was analyzed by 15% SDS-PAGE. The gel was stained with Coomassie Brilliant Blue; (B) The tRNA^Leu_UAG transcript was methylated by the Ta0836 gene product and then the generated methylated nucleotide was analyzed by 2D-TLC; (C) The methyl group acceptance activities of the wild-type tRNA^Leu_UAG transcript (lane 1) and the mutant tRNA^Leu_UAG transcript (lane 2), in which G37 was replaced by A, were investigated. The transcripts were individually incubated with the Ta0836 gene product and ¹⁴C-AdoMet, and then separated by 10% PAGE (7 M urea). The gel was stained with methylene blue (left panel) and the autoradiogram of the same gel was taken (right panel).