Enzymatic Synthesis of an RNA Fragment Corresponding to the Anti-Codon Loop and Stem of tRNAPhefrom Yeast

Abstract
A hexadecamer corresponding to the anticodon loop and stem of tRNAPhe yeast has been prepared using T4 RNA ligase and isolated by high performance liquid chromatography. The two oligonucleotides used in the ligation were isolated from a ribonuclease T1 digest of the tRNA which was resolved by HPLC on an anion exchange column. To prepare the “acceptor” oligonucleotide for the RNA ligase reaction a 3′ terminal phosphate was removed. To prepare the “donor” oligomer a 5′ terminal phosphate was added. Analysis of the product hexadecamer was by nucleoside and nucleotide-3′-mono-phosphate composition.