Automated chemical synthesis of biologically active tRNA having a sequence corresponding to Ascaris suum mitochondrial tRNA(Met) toward NMR measurements.
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.
Ohtsuki
T
T
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo.
Vinayak
R
R
Watanabe
Y
Y
Kita
K
K
Kawai
G
G
Watanabe
K
K
D000818:Animals; D017165:Ascaris suum; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*; D000077278:RNA, Mitochondrial; D012358:RNA, Transfer, Met / Q000737:chemistry*
D000077278:RNA, Mitochondrial; D012358:RNA, Transfer, Met; D012313:RNA