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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 Ta0931 gene product (Trm56) can methylate both the elongator tRNA^Met transcript and its precursor with an intron. (A) An aliquot of 4 µg of the Ta0931 gene product was analyzed by 15% SDS-PAGE. The gel was stained with Coomassie Brilliant Blue; (B) The Ta0931 gene product and tRNA^Leu_UAG transcript were incubated at 50 °C for 1 h, and then the ¹⁴C-methylated nucleotide was analyzed by 2D-TLC; (C) The incorporation of methyl groups into the mature transcript and the precursor of elongator tRNA^Met were investigated. The samples were as follows: lane 1, mature transcript of elongator tRNA^Met; lane 2, precursor of elongator tRNA^Met. The gel was stained with methylene blue (left panel) and the autoradiogram of the same gel was taken (right panel); (D) The incorporation of methyl groups into the mature transcript and the precursor of elongator tRNA^Met were measured by the filter assay. Closed and open circles show the incorporation of ¹⁴C-methyl groups into the mature transcript and precursor of elongator tRNA^Met, respectively.

The Ta0931 gene product (Trm56) can methylate both the elongator tRNA^Met transcript and its precursor with an intron. (A) An aliquot of 4 µg of the Ta0931 gene product was analyzed by 15% SDS-PAGE. The gel was stained with Coomassie Brilliant Blue; (B) The Ta0931 gene product and tRNA^Leu_UAG transcript were incubated at 50 °C for 1 h, and then the ¹⁴C-methylated nucleotide was analyzed by 2D-TLC; (C) The incorporation of methyl groups into the mature transcript and the precursor of elongator tRNA^Met were investigated. The samples were as follows: lane 1, mature transcript of elongator tRNA^Met; lane 2, precursor of elongator tRNA^Met. The gel was stained with methylene blue (left panel) and the autoradiogram of the same gel was taken (right panel); (D) The incorporation of methyl groups into the mature transcript and the precursor of elongator tRNA^Met were measured by the filter assay. Closed and open circles show the incorporation of ¹⁴C-methyl groups into the mature transcript and precursor of elongator tRNA^Met, respectively.