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. 2006 Jan 17;34(2):407–416. doi: 10.1093/nar/gkj445

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© The Author 2006. Published by Oxford University Press. All rights reserved

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Characterization of double mutant Est3p-D49A-K68A. (A) Telomere Southern blot. The isogenic strains are as following: wild-type (YPH499), D49A mutant, D49AK68A double mutant, K68A mutant. (B) Commassie blue staining of purified Est3p-D49A-K68A. Lane 1, uninduced total extract; lane 2, induced total extract; lane 3, GST-Est3p-D49A-K68A fusion protein on glutathione beads; lane 4, Est3p-D49A-K68A after thrombin digestion. (C) Analysis of the Est3p-D49A-K68A double mutant by gel filtration with superdex 75 column. The upper panel is the elution profile, and the lower panel is western blot using affinity purified polyclonal anti-Est3p antibodies.