Aminoacylation of tRNA by antisense molecule.
Aminoacylation of tRNA was attempted through formation of tRNA/DNA/aa-PNA (N-aminoacylated peptide nucleic acid) ternary hybrid. A 23-mer DNA, that is complementary to a 3'-terminal of tRNA and to a 9-mer PNA carrying an amino acid unit, was designed to achieve close proximity between the amino acid and the 3'-OH group of tRNA. The aminoacylation was carried out in a buffer solution containing imidazole. The aminoacylation was detected by nuclease S1 treatment followed by HPLC and MALDI-TOF MS. This novel methodology will open a way for easy and versatile aminoacylation of nonnatural amino acids onto specific tRNAs.
Ninomiya
Keiko
K
Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, Okayama, 700-8530, Japan.
Endo
Takamasa
T
Minohata
Toshikazu
T
Tajiri
Masahisa
M
Suzuki
Maya
M
Kurita
Tomoyoshi
T
Hohsaka
Takahiro
T
Sisido
Masahiko
M
D000215:Acylation; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012343:RNA, Transfer / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D002021:Buffers; D016376:Oligonucleotides, Antisense; D012343:RNA, Transfer