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October 1, 1981Proceedings of the National Academy of SciencesOpen Access

Molecular cloning of the genome of poliovirus type 1.

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Population

Mahoney strain of poliovirus type 1

Design

Preclinical

Authors

SWSylvie van der WerfFBFrançois BrégégèreHKH. Kopecká

Discussion

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Overview

May enable poliovirus molecular tools; leaves open clinical translation pending human validation.

Key Points

  • To clone the RNA genome of poliovirus type 1 into bacterial plasmids and construct a comprehensive physical restriction map.
  • Synthesized cDNA-RNA hybrids from the poliovirus type 1 Mahoney strain using reverse transcriptase and inserted them into Escherichia coli plasmid pBR322.
  • Identified recombinant colonies through in situ hybridization with virus-specific RNase T1-resistant oligonucleotides.
  • Characterized cloned cDNA inserts using restriction endonuclease mapping and electron microscopy.
  • Isolated overlapping cDNA clones up to 3.2 kilobase pairs in length, spanning at least 99% of the viral RNA genome.
  • Reconstructed the complete restriction map of the poliovirus genome from overlapping fragments.
  • Cloned the complete 5' end of the viral genome within recombinant plasmid pPV1-366.

Structured PICO

P
Population
Mahoney strain of poliovirus type 1
I
Intervention
Molecular cloning using reverse transcriptase and Escherichia coli plasmid pBR322
O
Outcome
Successful cloning and restriction mapping of the poliovirus genome

The study successfully cloned nearly the entire genome of poliovirus type 1, enabling detailed restriction mapping and further molecular analysis.

Cite This Study

Werf et al. (1981) studied this question.

synapsesocial.com/papers/6a722720b27f15817827f481https://doi.org/10.1073/pnas.78.10.5983
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Also Consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

  1. 1Sequence of 1060 3'-terminal nucleotides of poliovirus RNA as determined by a modification of the dideoxynucleotide method.1980 · 31 citations
  2. 2Sequence of picornavirus RNAs containing a radioiodinated 5'-linked peptide reveals a conserved 5' sequence.1980 · 32 citations
  3. 3Stable hairpin structure within the 5'-terminal 85 nucleotides of poliovirus RNA1981 · 57 citations
  4. 4Poliovirus Polyuridylic Acid Polymerase and RNA Replicase Have the Same Viral Polypeptide1979 · 112 citations
  5. 5A rapid alkaline extraction procedure for screening recombinant plasmid DNA1979 · 14,859 citations