Figure 8

Dot-blot analysis of total tRNA from the yfhQ K.O. strain incubated with YfhQ and AdoMet. The reaction mixture containing 50 mM PIPES-Na, pH 7.0, 4 mM MgCl₂, 100 μg of total tRNA from yfhQ K.O. strain, 5 μg of the purified YfhQ protein (1-YfhQ protein purified from the yfhQ K.O. strain transformed with a plasmid carrying the yfhQ gene, 2 – YfhQ purified from the regular overexpressing strain, 3 – a negative control experiment without any protein), and 10 μM [¹⁴C-methyl] AdoMet (53 mCi/mmol). After 60 min incubation at 37°C, 50 μl aliquots were filtered in duplicate in a dot-blot manifold (Scie-Plast) using a DE-81 cellulose filter. The filter was washed three times with 200 μl of the reaction buffer and one time with 70% EtOH, dried and exposed to the PhosphorImager screen. The resulting image was scanned on the Storm 820 PhosphorImager.