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PMID: 6158042 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Sequence of 1060 3'-terminal nucleotides of poliovirus RNA as determined by a modification of the dideoxynucleotide method.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 77 ·No. 6 ·1980-06-00 ·Pages 3196-200

Kitamura N, Wimmer E

Abstract

The dideoxynucleotide method for sequencing DNA developed by Sanger et al. [Sanger, F., Nicklen, S. & Coulson, A. (1977) Proc. Natl. Acad. Sci. USA 74, 5463-5467] was modified to allow sequence analysis of poliovirus RNA without recourse to cloning. Our method involves reverse transcription of poliovirus RNA followed by cDNA-dependent DNA synthesis in the presence of unlabeled dNTPs and 2',3'-dideoxynucleoside triphosphates, with Escherichia coli DNA polymerase I (Klenow) used to catalyze the reaction. DNA synthesis is primed by 5'-32P-labeled RNase T1- or RNase A-resistant oligonucleotides generated from poliovirus RNA. The sequence of 1060 nucleotides preceding the 3'-terminal poly(A) is presented. Based on the position of termination codons we propose that viral translation terminates at nucleotide -562.

MeSH Terms
Base Sequence Codon/analysis DNA/analysis DNA Polymerase I Electrophoresis, Polyacrylamide Gel Genes, Synthetic Poliovirus/genetics Protein Biosynthesis RNA, Viral/genetics RNA-Directed DNA Polymerase
Chemicals
Codon RNA, Viral DNA RNA-Directed DNA Polymerase DNA Polymerase I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kitamura N
Wimmer E
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38 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.77.6.3196
Published
1980-06-00
Pages
3196-200
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC349581
Subset
IM
Databases
GENBANK
J02281
External Links
PubMed source DOI: 10.1073/pnas.77.6.3196 PMC Full Text View journal details
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