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. 1977 Dec;4(12):4165–4174. doi: 10.1093/nar/4.12.4165

End labeling of enzymatically decapped mRNA.

A Efstratiadis, J N Vournakis, H Donis-Keller, G Chaconas, D K Dougall, F C Kafatos
PMCID: PMC343232  PMID: 202926

Abstract

A method is presented for rapid and efficient 5' end labeling with 32P of capped mRNAs, by a series of three enzymatic reactions: the blocking nucleotide of the cap structure is removed by tobacco acid pyrophosphatase, and after dephosphorylation with alkaline phosphatase the 5' end is labeled with gamma-32-P-ATP and T4 polynucleotide kinase.

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Selected References

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

  1. Abraham K. A., Lillehaug J. R. Enzymatic cleavage of m7 GDP from eucaryotic mRNA. FEBS Lett. 1976 Nov 15;72(1):49–52. doi: 10.1016/0014-5793(76)80896-4. [DOI] [PubMed] [Google Scholar]
  2. Abraham K. A., Pihl A. Translation of enzymically decapped messenger RNA. Eur J Biochem. 1977 Aug 1;77(3):589–593. doi: 10.1111/j.1432-1033.1977.tb11703.x. [DOI] [PubMed] [Google Scholar]
  3. Altenburg L. C., Getz M. J., Saunders G. F. 125I in molecular hybridization experiments. Methods Cell Biol. 1975;10:325–342. doi: 10.1016/s0091-679x(08)60744-x. [DOI] [PubMed] [Google Scholar]
  4. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Efstratiadis A., Kafatos F. C., Maniatis T. The primary structure of rabbit beta-globin mRNA as determined from cloned DNA. Cell. 1977 Apr;10(4):571–585. doi: 10.1016/0092-8674(77)90090-3. [DOI] [PubMed] [Google Scholar]
  6. Gamborg O. L. The effects of amino acids and ammonium on the growth of plant cells in suspension culture. Plant Physiol. 1970 Apr;45(4):372–375. doi: 10.1104/pp.45.4.372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gilbert W., Maxam A. The nucleotide sequence of the lac operator. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3581–3584. doi: 10.1073/pnas.70.12.3581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lillehaug J. R., Kleppe K. Effect of salts and polyamines on T4 polynucleotide kinase. Biochemistry. 1975 Mar 25;14(6):1225–1229. doi: 10.1021/bi00677a021. [DOI] [PubMed] [Google Scholar]
  9. Lockhard R. E., Rajbhandary U. L. Nucleotide sequences at the 5'termini of rabbit alpha and beta globin mRNA. Cell. 1976 Dec;9(4 Pt 2):747–760. doi: 10.1016/0092-8674(76)90138-0. [DOI] [PubMed] [Google Scholar]
  10. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ohno T., Okada Y., Shimotohno K., Miura K., Shinshi H. Enzymatic removal of the 5'-terminal methylated blocked structure of tobacco mosaic virus RNA and its effects on infectivity and reconstitution with coat protein. FEBS Lett. 1976 Aug 15;67(2):209–213. doi: 10.1016/0014-5793(76)80368-7. [DOI] [PubMed] [Google Scholar]
  12. Panet A., van de Sande J. H., Loewen P. C., Khorana H. G., Raae A. J., Lillehaug J. R., Kleppe K. Physical characterization and simultaneous purification of bacteriophage T4 induced polynucleotide kinase, polynucleotide ligase, and deoxyribonucleic acid polymerase. Biochemistry. 1973 Dec 4;12(25):5045–5050. doi: 10.1021/bi00749a003. [DOI] [PubMed] [Google Scholar]
  13. Scherberg N. H., Refetoff S. Radioiodine labeling of ribopolymers for special applications in biology. Methods Cell Biol. 1975;10:343–359. doi: 10.1016/s0091-679x(08)60745-1. [DOI] [PubMed] [Google Scholar]
  14. Shatkin A. J. Capping of eucaryotic mRNAs. Cell. 1976 Dec;9(4 Pt 2):645–653. doi: 10.1016/0092-8674(76)90128-8. [DOI] [PubMed] [Google Scholar]
  15. Shimotohno K., Kodama Y., Hashimoto J., Miura K. I. Importance of 5'-terminal blocking structure to stabilize mRNA in eukaryotic protein synthesis. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2734–2738. doi: 10.1073/pnas.74.7.2734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shinshi H., Miwa M., Kato K., Noguchi M., Matsushima T., Sugimura T. A novel phosphodiesterase from cultured tobacco cells. Biochemistry. 1976 May 18;15(10):2185–2190. doi: 10.1021/bi00655a024. [DOI] [PubMed] [Google Scholar]
  17. Shinshi H., Miwa M., Sugimura T. Enzyme cleaving the 5'-terminal methylated blocked structure of messenger RNA. FEBS Lett. 1976 Jun 1;65(2):254–257. doi: 10.1016/0014-5793(76)80492-9. [DOI] [PubMed] [Google Scholar]
  18. Weiss B., Live T. R., Richardson C. C. Enzymatic breakage and joining of deoxyribonucleic acid. V. End group labeling and analysis of deoxyribonucleic acid containing single straned breaks. J Biol Chem. 1968 Sep 10;243(17):4530–4542. [PubMed] [Google Scholar]
  19. Wimmer E. Sequence studies of poliovirus RNA. I. Characterization of the 5'-terminus. J Mol Biol. 1972 Jul 28;68(3):537–540. doi: 10.1016/0022-2836(72)90106-4. [DOI] [PubMed] [Google Scholar]

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