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. 2012 Jun 28;7(6):e39297. doi: 10.1371/journal.pone.0039297

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This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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Mutants, K225A, Y242A, F248A, E265A, and D293A were introduced into the hTYW2 gene by oligo-directed mutagenesis. These mutant protein expression constructs in the pYES2 vector were used to transform the yeast TYW2 deletion strain (ΔTYW2). All the mutant constructs expressed the TYW2 protein (not shown). LC/MS analysis of nuclease P1 digested tRNA-Phe obtained from the deletion strain ΔTYW2 transformed with wild type (top left) and the five mutant hTYW2 constructs in pYES2. For each transformant, the panels show UV at 254 nm (top), and mass chromatograms detecting MH+ (m/z838) of yWpA (middle), and MH+ (m/z 651) of [yW-187]pA (bottom) respectively.