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.
AUTOR(ES)
Simoncsits, A
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=324222Documentos Relacionados
- Complete nucleotide sequence of a 16S ribosomal RNA gene from Streptomyces griseus subsp. griseus.
- Isolation of Uridine 5′-Pyrophosphate Glucuronic Acid Pyrophosphorylase and Its Assay Using 32P-Pyrophosphate 1
- Nucleotide sequence of the streptomycinphosphotransferase and amidinotransferase genes from Streptomyces griseus.
- Nucleotide sequence analysis of precursor 5S RNA from Bacillus licheniformis.
- Cleavage of adenosine 5'-monophosphate during uptake by Streptomyces griseus.
