title: The preparative synthesis of oligodeoxyribonucleotides using RNA ligase.
abstract: The synthesis of nmol quantities of defined sequences of oligodeoxyribonucleotides using T4 RNA ligase has been demonstrated. Reacting using from 18 to 200 nmol of substrates in which a single 2'-deoxyribonucleoside 3',5'-bisphosphate was added to an oligodeoxyribonucleotide resulted in yields from 13 to 95%. When two oligodeoxyribonucleotides were similarly joined using RNA ligase, the yields ranged from 10 to 50%. Although the reactions contained high concentrations of enzyme and were incubated from 5 to 21 days, there was little degradation of either substrates or products. We have also characterized an unusual product which arises when 3'-phosphate terminated oligodeoxyribonucleotides are incubated with RNA ligase and high concentrations of ATP. This product has an adenylyl group linked to the 3'-phosphate by an anhydride bond. The mechanistic and synthetic implications of forming this product are discussed.
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology; D008722:Methods; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis*; D009841:Oligonucleotides / Q000138:chemical synthesis*; D010761:Phosphorus Radioisotopes; D011118:Polynucleotide Ligases / Q000378:metabolism*; D011130:RNA Ligase (ATP) / Q000378:metabolism*; D013379:Substrate Specificity; D013604:T-Phages / Q000201:enzymology
chemical_list: D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D011118:Polynucleotide Ligases; D011130:RNA Ligase (ATP)