PMID: 6986903Feb 19, 1980Paper

T4 ribonucleic acid ligase joins single-strand oligo(deoxyribonucleotides)

Biochemistry
M I McCoy, R I Gumport

Abstract

T4 RNA ligase joins a 3'-hydroxyl-terminated acceptor oligoribonucleotide to a 5'-phosphate-terminated donor oligoribonucleotide. An analogous reaction with single-strand DNA oligonucleotides would be useful for the synthesis of defined sequences of DNA because it would eliminate the need to synthesize complementary sequences to form the duplex substrates required by DNA ligase. We have studied the model reaction dA(pdA)5 + [5'-32P] (pdT)4pdCp leads to dA(pdA)5 [3' leads to 5'-32P]pdT(pdT)3pdCp and have obtained 40-60% yields at equimolar concentrations (100 microM to 1 mM) of the two substrates. Higher yields have been obtained when acceptor concentrations in excess of those of the donor are used. The use of a 5'-hydroxyl, 3'-hydroxyl terminated acceptor and a 5'-phosphate, 3'-phosphate terminated donor limits the reaction to a unique product. The 3'-phosphate-terminated donor was prepared by using RNA ligase to add a single deoxyribonucleoside 3',5'-bisphosphate donor to an oligo(deoxyribonucleotide) acceptor [Hinton, D.M., Baez, J.A., & Gumport, R.I. (1978) Biochemistry 17, 5091]. The DNA oligomer joining reaction requires low concentrations of ATP and an ATP regenerating system, Mn2+, high levels of nuclease-free RNA ligase...Continue Reading

References

Jan 1, 1977·Nucleic Acids Research·O C Uhlenbeck, V Cameron
Jul 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·B MeyhackN R Pace
Mar 1, 1978·Nucleic Acids Research·V CameronO C Uhlenbeck
Mar 28, 1979·Biochimica Et Biophysica Acta·M I McCoyR I Gumport
Mar 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·Y KikuchiK Sakaguchi
Sep 25, 1979·Nucleic Acids Research·D M Hinton, R I Gumport
Feb 1, 1978·Nucleic Acids Research·Y Kikuchi, K Sakaguchi
May 30, 1978·Biochemistry·T E England, O C Uhlenbeck
Jan 26, 1976·Biochemical and Biophysical Research Communications·T J SnopekN R Cozzarelli
Jan 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·G C WalkerR I Gumport
Oct 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·R SilberJ Hurwitz
Sep 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·G Kaufmann, U Z Littauer
Jun 1, 1977·Proceedings of the National Academy of Sciences of the United States of America

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Citations

Nov 13, 1991·Biochimica Et Biophysica Acta·A R Beaudoin, G Grondin
Aug 24, 2000·Proceedings of the National Academy of Sciences of the United States of America·R A Lease, M Belfort
Feb 26, 2010·Nucleosides, Nucleotides & Nucleic Acids·Anilkumar R KoreScott Kuersten
Mar 25, 1982·Nucleic Acids Research·D M HintonR I Gumport
Dec 5, 2003·Nucleic Acids Research·Thorarinn BlondalJakob K Kristjansson
Nov 8, 2013·Nucleic Acids Research·Carsten A RaabeTimofey S Rozhdestvensky
Jan 15, 1982·Journal of Molecular Biology·J M RunnelsL Snyder
Oct 1, 1986·Analytical Biochemistry·D C TessierT Vernet
Jan 15, 1985·Biochemistry·L W McLaughlinE Graeser
Apr 4, 2017·Nucleic Acids Research·Christos ArgyropoulosDavid Galas
May 25, 2021·Computational and Structural Biotechnology Journal·Eojin YooByung-Kwan Cho

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