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. 1977 Oct;4(10):3341–3355. doi: 10.1093/nar/4.10.3341

A simple method for the preparation of [beta-32P]purine nucleoside triphosphase.

Y Furuichi, A J Shatkin
PMCID: PMC342657  PMID: 928062

Abstract

A rapid, simple and inexpensive procedure is described for the preparation of purine ribo-and deoxyribonucleoside triphosphates specifically and highly radiolabeled with [32P]phosphate in the beta position. The method involves two successive enzymatic reactions: conversion of donor [gamma-32P]ATP in the presence of an excess of acceptor 5'-mononucleotide to the 5'-diphosphates by myokinase or guanosine 5'-monophosphate kinase followed by phosphorylation with pyruvate kinase to yield 5'-triphosphates.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams J. M., Cory S. Modified nucleosides and bizarre 5'-termini in mouse myeloma mRNA. Nature. 1975 May 1;255(5503):28–33. doi: 10.1038/255028a0. [DOI] [PubMed] [Google Scholar]
  2. Borsa J., Grover J., Chapman J. D. Presence of nucleoside triphosphate phosphohydrolase activity in purified virions of reovirus. J Virol. 1970 Sep;6(3):295–302. doi: 10.1128/jvi.6.3.295-302.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dubin D. T., Taylor R. H. The methylation state of poly A-containing messenger RNA from cultured hamster cells. Nucleic Acids Res. 1975 Oct;2(10):1653–1668. doi: 10.1093/nar/2.10.1653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Furuichi Y., Morgan M., Muthukrishnan S., Shatkin A. J. Reovirus messenger RNA contains a methylated, blocked 5'-terminal structure: m-7G(5')ppp(5')G-MpCp-. Proc Natl Acad Sci U S A. 1975 Jan;72(1):362–366. doi: 10.1073/pnas.72.1.362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Furuichi Y., Morgan M., Shatkin A. J., Jelinek W., Salditt-Georgieff M., Darnell J. E. Methylated, blocked 5 termini in HeLa cell mRNA. Proc Natl Acad Sci U S A. 1975 May;72(5):1904–1908. doi: 10.1073/pnas.72.5.1904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Furuichi Y., Muthukrishnan S., Tomasz J., Shatkin A. J. Mechanism of formation of reovirus mRNA 5'-terminal blocked and methylated sequence, m7GpppGmpC. J Biol Chem. 1976 Aug 25;251(16):5043–5053. [PubMed] [Google Scholar]
  7. Furuichi Y., Shatkin A. J. 5'-termini of reovirus mRNA: ability of viral cores to form caps post-transcriptionally. Virology. 1977 Apr;77(2):566–578. doi: 10.1016/0042-6822(77)90482-2. [DOI] [PubMed] [Google Scholar]
  8. Furuichi Y., Shatkin A. J. Differential synthesis of blocked and unblocked 5'-termini in reovirus mRNA: effect of pyrophosphate and pyrophosphatase. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3448–3452. doi: 10.1073/pnas.73.10.3448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gissinger F., Kedinger C., Chambon P. Animal DNA-dependent RNA polymerases. 10. General enzymatic properties of purified calf thymus RNA polymerases AI and B. Biochimie. 1974;56(3):319–333. doi: 10.1016/s0300-9084(74)80139-2. [DOI] [PubMed] [Google Scholar]
  10. Glynn I. M., Chappell J. B. A simple method for the preparation of 32-P-labelled adenosine triphosphate of high specific activity. Biochem J. 1964 Jan;90(1):147–149. doi: 10.1042/bj0900147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kapuler A. M., Mendelsohn N., Klett H., Acs G. Four base-specific nucleoside 5'-triphosphatases in the subviral core of reovirus. Nature. 1970 Mar 28;225(5239):1209–1213. doi: 10.1038/2251209a0. [DOI] [PubMed] [Google Scholar]
  12. Karstadt M., Krakow J. S. Azotobacter vinelandii ribonucleic acid polymerase. IX. Alternating copolymer of inosinic and cytidylic residues-directed synthesis of alternating copolymer of guanylic and cytidylic residues. J Biol Chem. 1970 Feb 25;245(4):746–751. [PubMed] [Google Scholar]
  13. Kates J., Beeson J. Ribonucleic acid synthesis in vaccinia virus. I. The mechanism of synthesis and release of RNA in vaccinia cores. J Mol Biol. 1970 May 28;50(1):1–18. doi: 10.1016/0022-2836(70)90100-2. [DOI] [PubMed] [Google Scholar]
  14. LITTAUER U. Z., KIMHI Y., AVRON M. A METHOD FOR THE SYNTHESIS OF LABELED RIBONUCLEOTIDES. Anal Biochem. 1964 Sep;9:85–93. doi: 10.1016/0003-2697(64)90086-7. [DOI] [PubMed] [Google Scholar]
  15. Lavi S., Shatkin A. J. Methylated simian virus 40-specific RNA from nuclei and cytoplasm of infected BSC-1 cells. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2012–2016. doi: 10.1073/pnas.72.6.2012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moss B., Gershowitz A., Wei C. M., Boone R. Formation of the guanylylated and methylated 5'-terminus of vaccinia virus mRNA. Virology. 1976 Jul 15;72(2):341–351. doi: 10.1016/0042-6822(76)90163-x. [DOI] [PubMed] [Google Scholar]
  17. Paolette E., Rosemond-Hornbeak H., Moss B. Two nucleid acid-dependent nucleoside triphosphate phosphohydrolases from vaccinia virus. Purification and characterization. J Biol Chem. 1974 May 25;249(10):3273–3280. [PubMed] [Google Scholar]
  18. Perry R. P., Kelley D. E., Friderici K., Rottman F. The methylated constituents of L cell messenger RNA: evidence for an unusual cluster at the 5' terminus. Cell. 1975 Apr;4(4):387–394. doi: 10.1016/0092-8674(75)90159-2. [DOI] [PubMed] [Google Scholar]
  19. Raué H. A., Cashel M. Preparation of (beta-32P)ribonucleoside-5'-triposphates using permeable cells of Escherichia coli. Anal Biochem. 1973 Nov;56(1):129–136. doi: 10.1016/0003-2697(73)90176-0. [DOI] [PubMed] [Google Scholar]
  20. Reichard P., Eliasson R., Söderman G. Initiator RNA in discontinuous polyoma DNA synthesis. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4901–4905. doi: 10.1073/pnas.71.12.4901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Richardson J. P. RNA polymerase and the control of RNA synthesis. Prog Nucleic Acid Res Mol Biol. 1969;9:75–116. doi: 10.1016/s0079-6603(08)60768-0. [DOI] [PubMed] [Google Scholar]
  22. SINGER M. F., GUSS J. K. The dependence of reactions catalyzed by polynucleotide phosphorylase on oligonucleotides. J Biol Chem. 1962 Jan;237:182–189. [PubMed] [Google Scholar]
  23. Shatkin A. J., LaFiandra A. J. Transcription by infectious subviral particles of reovirus. J Virol. 1972 Oct;10(4):698–706. doi: 10.1128/jvi.10.4.698-706.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Warner A. H. A simple and inexpensive method for the preparation of (beta,gamma-32P) guanosine 5'-triphosphate. Biochim Biophys Acta. 1975 Mar 21;383(3):229–235. [PubMed] [Google Scholar]
  25. Wei C. M., Gershowitz A., Moss B. Methylated nucleotides block 5' terminus of HeLa cell messenger RNA. Cell. 1975 Apr;4(4):379–386. doi: 10.1016/0092-8674(75)90158-0. [DOI] [PubMed] [Google Scholar]
  26. Weissbach A. Vertebrate DNA polymerases. Cell. 1975 Jun;5(2):101–108. doi: 10.1016/0092-8674(75)90017-3. [DOI] [PubMed] [Google Scholar]

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