T4 RNA ligase catalyzes the ATP-dependent phosphodiester bond formation between a 5´-terminal phosphate (donor) and a 3´-terminal hydroxyl (acceptor). Although the enzyme can produce circular products with substrates of sufficient length that contain the required termini RNA ligase is also a very effective intermolecular joining reagent for oligonucleotide synthesis. The versatility of RNA ligase as a reagent for oligoribonucleotide synthesis is illustrated by the following two examples. For preparative synthesis, the enzyme readily catalyzes the equimolar joining of two oligoribonucleotide blocks on a moderate scale (ca. 1 μmol) and the enzyme can be used to synthesize small amounts of internally P-labeled oligomers of high specific activity. This chapter discusses applications for oligoribonucleotide synthesis and also explains the use of RNA ligase for the preparation of deoxy oligomers.
ASJC Scopus subject areas
- Molecular Biology