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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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T. acidophilum Trm1 can methylate both the elongator tRNA^Met transcript and its precursor, which contains an intron. The mature transcript of elongator tRNA^Met and its precursor with an intron (Figure 1D) were prepared. (A) These transcripts were incubated with T. acidophilum Trm1 and ¹⁴C-AdoMet at 50 °C for 5 min, and then separated by 10% PAGE (7 M urea) (left panel). The gel was stained with methylene blue. The samples are as follows: lane 1, mature transcript of elongator tRNA^Met; lane 2, precursor of elongator tRNA^Met with an intron. The right panel shows an autoradiogram of the same gel; (B) The ¹⁴C-methylated nucleosides in the mature transcript of elongator tRNA^Met were analyzed by 2D-TLC.