PMID: 6159585Sep 25, 1980Paper

3' Terminal labelling of RNA of RNA with beta-32P-pyrophosphate group and its application to the sequence analysis of 5S RNA from Streptomyces griseus

Nucleic Acids Research
A Simoncsits

Abstract

Nucleotide pyrophosphate transferase isolated from Streptomyces griseus is used to transfer pyrophosphate group from gamma-32P-ATP to the 3'-OH of tRNA, generating a strictly terminal label at its 3' end. Using yeast tRNAPhe as model compound, it is demonstrated that the labelled molecule is suitable for rapid gel sequencing by both enzymatic and chemical methods. RNA molecules terminated by pyrimidine nucleoside are poor pyrophosphate acceptors. To label RNAs of this kind, first guanosine 5'-phosphate 3'-(beta-32P)-pyrophosphate (pGpp) is prepared from gamma-32P-ATP and GMP by nucleotide pyrophosphate transferase. pGpp is then ligated to the 3' end of RNA by T4 RNA ligase. The complete nucleotide sequence of 5S RNA from Streptomyces griseus is established by rapid gel sequencing methods performed on 3'-(beta-32P)-pyrophosphate labelled molecule.

References

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Citations

Feb 1, 1988·Molecular & General Genetics : MGG·H A Baylis, M J Bibb
Dec 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·D A PeattieH F Noller
Jan 11, 1982·Nucleic Acids Research·D SankoffW McKay
May 11, 1982·Nucleic Acids Research·R M MacKayW F Doolittle
Jan 11, 1983·Nucleic Acids Research·V A ErdmannR De Wachter
Nov 11, 1983·Nucleic Acids Research·H NeimarkN Delihas
Feb 11, 1983·Nucleic Acids Research·H KüntzelU Hahn
Jan 1, 1978·Nucleic Acids Research·V A Erdmann
Jan 1, 1987·Journal of Molecular Evolution·E DamsR De Wachter
Jun 1, 1993·Zentralblatt Für Bakteriologie : International Journal of Medical Microbiology·Y H ParkM Goodfellow
May 1, 1987·Journal of Bacteriology·Y H ParkK Komagata
Jul 1, 1984·Journal of Bacteriology·S DekioS Osawa

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