Enzymatic Synthesis of Oligoribonucleotide

Abstract

In order to systematically synthesize oligoribonucleotides with defined sequence, trimer unit having different bloking groups at 3'-hydroxyl and 5'-phosphate groups were enzymatically synthesizd. Deblocking at 3'- or 5'-end of the trimer unit gives an acceptor or a donor for the T4RNA ligase, respectively. For the blocking of the 3'-end, o-nitrobenzyl group was attached to the 2'-hydroxyl group, while for the blocking of 5'-phosphate, anilidate was employed. The f ormer was deblocked by photo-irradiation and the latter was by an acid. The hexamer produced by the ligation of the acceptor and the donoralso has the same blocking groups at 3'- and 5'-ends, and is capable to be utilized for the next synthetic reaction as an acceptor or a donor for T4RNA ligase.