The synthesis of oligodeoxyribonucleotides using RNA ligase
- 1 January 1979
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 7 (2) , 453-464
- https://doi.org/10.1093/nar/7.2.453
Abstract
T4 RNA ligase catalyzes the addition of a single deoxyribonucleoside 3',5'-bisphosphate to the 3'-hydroxyl of oligodeoxyribonucleotides (Hinton et al. (1978) Biochemistry 17, 5091). We have determined improved conditions for this reaction which give yields equal to or greater than 85% when any of five common deoxyribonucleoside bisphosphate (pdAp, pdCp, pdGp, pdTp, or pdUp) are added to dA(PDA)4. A low ATP concentration, which is constantly maintained by a regeneration system composed of phosphocreatine, creatine kinase, and myokinase, contributes to the attainment of high yields. The addition of RNase A and spermine also enhances the rates and yields of the reactions. These conditions facilitate the use of RNA ligase as a reagent for the stepwise synthesis of DNA of defined sequence.Keywords
This publication has 17 references indexed in Scilit:
- The purification of nuclease-free T4-RNA ligaseBiochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1979
- T4 RNA ligase joins 2'-deoxyribonucleoside 3',5'-bisphosphates to oligodeoxyribonucleotidesBiochemistry, 1978
- Reactions at the termini of tRNA with T4 RNA ligaseNucleic Acids Research, 1978
- Synthesis of modified nucleoside 3',5'-bisphosphates and their incorporation into oligoribonucleotides with T4 RNA ligaseBiochemistry, 1978
- Enzymic oligoribonucleotide synthesis with T4 RNA ligaseBiochemistry, 1978
- Polynucleotide kinase from a T4 mutant which lacks the 3′ phosphatase activityNucleic Acids Research, 1978
- Equimolar addition of oligoribonucleotides with T4 RNA ligaseNucleic Acids Research, 1977
- The use of terminal blocking groups for the specific joining of oligonucleotides in RNA ligase reactions containing equimolar concentrations of acceptor and donor moleculesNucleic Acids Research, 1976
- Some effects of charge and structure upon ionic interactions of nucleic acidsBiochimica et Biophysica Acta, 1961
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951