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PMID: 229466 Published · ppublish English Journal Article

A convenient sequencing method for 5' protein-linked RNAs.

Nucleic acids research ·Vol. 7 ·No. 5 ·1979-11-10 ·Pages 1233-46

Nomoto A, Imura N

Abstract

A convenient nucleotide sequencing method for 5' end protein-linked RNAs was developed. Genome of LSc, 2ab poliovirus, which has a protein (VPg) covalently linked to the 5' terminus, was labelled with 125I Bolton and Hunter reagent after proteinase K treatment. No sign of labelling of nucleotide moiety in the genome with the reagent was detected. A labelled oligo peptide-linked ribonuclease T1 fragment was obtained from the 5' end of the genome. Analysis of the complex by two dimensional gel electrophoresis after partial alkali digestion or by the nucleotide sequencing method of Donis-Keller et al. (1) revealed that LSc, 2ab strain genome had an identical 5' end structure to that of Mahoney strain genome, that is, VPg-pUpUpApApApApCpApGp. Our results have shown that this labelling method is useful for analysis of 5' end sequence of RNAs linked to protein at the 5' termini.

MeSH Terms
Base Sequence Genes, Viral Methods Nucleoproteins Poliovirus/analysis RNA, Viral Ribonuclease T1 Ribonucleases Ribonucleoproteins
Chemicals
Nucleoproteins RNA, Viral Ribonucleoproteins Ribonucleases Ribonuclease T1
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Nomoto A
Imura N
References (22)
22 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/7.5.1233
Published
1979-11-10
Pages
1233-46
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342299
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/7.5.1233 PMC Full Text View journal details
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