Aminoacyl-tRNA synthesis by a resin-immobilized ribozyme

J Am Chem Soc. 2002 Jun 19;124(24):6834-5. doi: 10.1021/ja025872a.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biotin / analogs & derivatives
  • Enzymes, Immobilized / chemistry*
  • Enzymes, Immobilized / metabolism
  • Nucleic Acid Conformation
  • Phenylalanine / analogs & derivatives
  • RNA, Catalytic / chemistry*
  • RNA, Catalytic / metabolism
  • RNA, Transfer, Amino Acyl / chemical synthesis*
  • RNA, Transfer, Amino Acyl / isolation & purification
  • Sepharose / chemistry

Substances

  • Enzymes, Immobilized
  • RNA, Catalytic
  • RNA, Transfer, Amino Acyl
  • Phenylalanine
  • Biotin
  • Sepharose