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Nucleic Acids Research, 1989, Vol. 17, No. 7 2623-2638
© 1989

CHEMISTRY

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

Takashi Yokogawa, Yoshinori Kumazawa, Kin-chiro Miura and Kimitsuna Watanabe*,+

Department of Industrial Chemistry, Faculty of Engineering, University of Tokyo Bunkyo-ku, 113 Tokyo, Japan

*To whom correspondence should be addressed

Received January 3, 1989. Revised March 3, 1989. Accepted March 3, 1989.

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 (Formula and Formula) from bovine mitochondria. It is about 100 times more sensitive than the conventional aminoacylation assay method. 2-4 A260 units each of both tRNASer 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 Formula possesses the D loop/T loop interaction like usual L-shaped tRNAs, and that Formula lacking almost an entire D arm is aminoacylated with an efficiency not very much lower than that of Formula.

+Present address Department of Life Chemistry, Tokyo Institute of Technology, Nagatuta, Yokohama 227, Japan


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