Purification and characterization of two serine isoacceptor tRNAs from bovine mitochondria by using a hybridization assay method

Abstract

For large scale preparation of mitochondrial tRNAs, a new hybridization assay method using synthetic oligodeoxy-ribonucleotide probes (16–17mer) complementary to individual tRNA sequences was developed and applied for the purification of two serine isoacceptor tRNAs ( and ) from bovine mitochondria. It is about 100 times more sensitive than the conventional aminoacylation assay method. 2-4 A₂₆₀ units each of both tRNA^Ser isoacceptors were purified from 17.5 kg of bovine liver, and they were characterized by means of nuclease digestion, melting profiles and aminoacylation activity. It is suggested that possesses the D loop/T loop interaction like usual L-shaped tRNAs, and that lacking almost an entire D arm is aminoacylated with an efficiency not very much lower than that of .