<i>In vivo</i> Analysis of Mtr4p-K904N Mutant Expressing on High Copy Plasmid in <i>trm6-504</i>.

<p>A) Growth of <i>trm6-504/MTR4</i> transformant bearing high-copy plasmid-born <i>(</i>HC) wild-type <i>MTR4</i> or HC <i>mtr4-904</i>, by a serial dilution spot assay and incubation on SC-Ura at 30°C or 36°C. Growth of WT yeast strain <i>TRM6/MTR4</i> was used as a positive control. B) Northern blot of total RNA to detect tRNA_i^Met steady-state levels in <i>trm6-504</i> strains bearing HC <i>MTR4</i>-WT, or HC<i>mtr4</i>-<i>904</i>. 5S rRNA was detected and used to normalize the amount of tRNA_i^Met in each lane, and the percentage of tRNA_i^Met from each strain is shown relative to <i>trm6-504</i>. C) Western blot analysis of total protein to detect Mtr4p in <i>trm6-504</i> strain with HC <i>MTR4</i>, or HC <i>mtr4-904</i>, and Nab2p is detected as a loading control.