Automated Chemical Synthesis of Biologically Active tRNA Having a Sequence Corresponding to Ascaris suum Mitochondrial tRNAMet toward NMR Measurements

Takashi Ohtsuki; Ravi Vinayak; Yoh-ichi Watanabe; Kiyoshi Kita; Gota Kawai; Kimitsuna Watanabe

Automated Chemical Synthesis of Biologically Active tRNA Having a Sequence Corresponding to Ascaris suum Mitochondrial tRNAMet toward NMR Measurements

Takashi Ohtsuki, Ravi Vinayak, Yoh-ichi Watanabe, Kiyoshi Kita, Gota Kawai, Kimitsuna Watanabe

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Keywords: Ascaris suum, chemical synthesis, mitochondria, NMR, tRNA

JOURNAL FREE ACCESS

1996 Volume 120 Issue 6 Pages 1070-1073

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Abstract

RNA samples corresponding to Ascaris suum mitochondrial tRNA^Met were chemically and automatically synthesized in amounts sufficient for NMR measurement. Conventional and rapid deprotection methods gave tRNA samples with the same amino acid-accepting activity as those prepared by other method; enzymatic synthesis, and enzymatic ligation of chemically synthesized fragments. The synthetic tRNA showed the same ¹H-NMR spectrum in the iminoproton region as the ligated tRNA. This rapid and reliable preparation method thus provides biologically active tRNA for NMR measurement, and further, it is applicable for synthesis of other large synthetic RNAs, by combining the site-specific isotopic labeling method.

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