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PMID: 17097 Published · ppublish English Journal Article

Equimolar addition of oligoribonucleotides with T4 RNA ligase.

Nucleic acids research ·Vol. 4 ·No. 1 ·1977-01-00 ·Pages 85-98

Uhlenbeck OC, Cameron V

Abstract

T4 induced RNA ligase will join equimolar concentrations of two oligoribonucleotides, (Ap)3C and p(Up) 5, to form a single product, (Ap)3Cp(Up) 5, in high yield. The presence of the 3' phosphate on p(Up)5 prevents the oligomer from adding to itself. The pH optimum of the reaction is about 7.5, but less of the undesirable adenylated intermediate, App(Up) 5, forms at pH 8.2. The reaction rate is a linear function of oligomer concentration from 3 micronM to 0.6 mM. The data suggest that T4 RNA ligase will be a useful enzyme for the synthesis of oligomers of defined sequence.

MeSH Terms
Adenosine Triphosphate/pharmacology Coliphages/enzymology Dithiothreitol/pharmacology Escherichia coli Hydrogen-Ion Concentration Kinetics Mutation Oligonucleotides/metabolism Oligoribonucleotides/metabolism Polynucleotide Ligases/metabolism
Chemicals
Oligonucleotides Oligoribonucleotides Adenosine Triphosphate Polynucleotide Ligases Dithiothreitol
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Uhlenbeck O C
Cameron V
References (7)
7 references, click to expand
  1. T4-induced RNA ligase joins single-stranded oligoribonucleotides.
    Proc Natl Acad Sci U S A. 1975 Jan;72(1):122-6 PMID: 1090929
  2. Determination of recognition sites of T4 RNA ligase on the 3'-OH and 5' -P termini of polyribonucleotide chains.
    Nature. 1975 Apr 3;254(5499):452-4 PMID: 1118037
  3. Joining of ribooligonucleotides with T4 RNA ligase and identification of the oligonucleotide-adenylate intermediate.
    Nucleic Acids Res. 1976 Jun;3(6):1613-23 PMID: 183186
  4. Studies on ribonucleic acid ligase. Characterization of an adenosine triphosphate-inorganic pyrophosphate exchange reaction and demonstration of an enzyme-adenylate complex with T4 bacteriophage-induced enzyme.
    J Biol Chem. 1974 Dec 10;249(23):7447-56 PMID: 4373468
  5. DNA ligase: structure, mechanism, and function.
    Science. 1974 Nov 29;186(4166):790-7 PMID: 4377758
  6. Complementary oligonucleotide binding to the anticodon loop of fMet-transfer RNA.
    Nature. 1970 Feb 7;225(5232):508-10 PMID: 5411856
  7. Purification and properties of bacteriophage T4-induced RNA ligase.
    Arch Biochem Biophys. 1976 May;174(1):167-76 PMID: 938049
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1977-01-00
Pages
85-98
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342411
Subset
IM
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