Aminoacyl-tRNA synthesis by a resin-immobilized ribozyme.
We report herein a new method for the aminoacylation of tRNA, using a resin-immobilized ribozyme and the cyanomethyl ester (CME) of an amino acid substrate. The oxidized form of the ribozyme was immobilized on a hydrazine resin via covalent linkage. We performed aminoacylation of tRNAs using this ribozyme-resin to isolate aminoacyl-tRNAs. The column was recycled up to 5 times without significant activity loss. Thus, our ribozyme-based aminoacylation system has significant potential to be a powerful and practical technique for supplying various nonnatural aminoacyl-tRNAs for a highly efficient in vitro translation system.
Murakami
Hiroshi
H
Department of Chemistry, University at Buffalo, State University of New York, Buffalo, New York 14260-3000.
Bonzagni
Neil J
NJ
Suga
Hiroaki
H
D001710:Biotin / Q000031:analogs & derivatives; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D010649:Phenylalanine / Q000031:analogs & derivatives; D016337:RNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000138:chemical synthesis* / Q000302:isolation & purification; D012685:Sepharose / Q000737:chemistry
D004800:Enzymes, Immobilized; D016337:RNA, Catalytic; D012346:RNA, Transfer, Amino Acyl; D010649:Phenylalanine; D001710:Biotin; D012685:Sepharose