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. 2016 Aug;22(8):1239–1249. doi: 10.1261/rna.056077.116

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© 2016 Chung et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 7. — Insertion of an active site histidine into Gld2 alters nucleotide specificity. (A) Gld2 and Gld2-His purification. Recombinant His-tagged Gld2 and the Gld2-His mutant were purified via Ni-NTA chromatography to apparent homogeneity as judged by SDS-PAGE. (B,C) Nucleotide preference of Gld2 and Gld2-His. Recombinant enzymes were incubated with and without RNA substrates and radiolabeled ATP or UTP. Formation of α-³²P labeled RNA products was monitored by electrophoretic separation and subsequent phosphorimaging. RNA substrates were let-7a (B) and 15A (C).