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PMID: 6273836 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Molecular cloning of the genome of poliovirus type 1.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 78 ·No. 10 ·1981-10-00 ·Pages 5983-7

van der Werf S, Bregegere F, Kopecka H, Kitamura N, Rothberg PG, Kourilsky P, Wimmer E, Girard M

Abstract

Poliovirus cDNA.RNA hybrids were prepared from the Mahoney strain of poliovirus type 1 by using reverse transcriptase (RNA-dependent DNA nucleotidyltransferase) and cloned in the Escherichia coli plasmid pBR322. Bacteria colonies carrying recombinant plasmids were selected by in situ hybridization with virus-specific RNase T1-resistant oligonucleotides. Analysis of the cDNA inserts by restriction mapping and electron microscopy showed that the cloned cDNAs, the longest of which was 3.2 kilobase pairs, originated from various parts of the viral RNA, covering at least 99% of the genome length. Due to overlapping of the clones, the restriction map of the poliovirus genome could be reconstructed. The complete 5' end of the genome was successfully cloned in at least one of the recombinant plasmids, pPV1-366.

MeSH Terms

Cloning, Molecular DNA Restriction Enzymes DNA, Recombinant/metabolism Genes, Viral Nucleic Acid Hybridization Oligoribonucleotides/analysis Poliovirus/genetics Ribonuclease T1

Chemicals

DNA, Recombinant Oligoribonucleotides DNA Restriction Enzymes Ribonuclease T1

Authors & Affiliations

8 authors, click to expand affiliations / ORCID

van der Werf S

Bregegere F

Kopecka H

Kitamura N

Rothberg P G

Kourilsky P

Wimmer E

Girard M

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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.78.10.5983

Published

1981-10-00

Pages

5983-7

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC348961

Subset

Grants

PHS HHS · A1 15122 · United States

NCI NIH HHS · CA 28146 · United States

External Links

PubMed source DOI: 10.1073/pnas.78.10.5983 PMC Full Text View journal details

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Molecular cloning of the genome of poliovirus type 1.

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