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The Journal of general virology1981; 57(Pt 2); 307-323; doi: 10.1099/0022-1317-57-2-307

Analysis of the genome of equine herpesvirus type 1: arrangement of cleavage sites for restriction endonucleases EcoRI, BglII and BamHI.

Abstract: The genome of an Australian isolate of equine herpesvirus type 1 (equine abortion virus) has been analysed using the restriction endonucleases EcoRI, BglII and BamHI, and a physical map constructed. Terminal fragments were identified by exonuclease treatments, and linkage of fragments was deduced by a combination of single- and double-digest experiments and cross-blot hybridizations. The genome has a mol. wt. of 100 x 10(6) and is comprised of a short unique region bounded by repetitive sequences, which is present in both orientations in approximately equal amounts in the DNA population, and a long unique region existing in only one orientation.
Publication Date: 1981-12-01 PubMed ID: 6275028DOI: 10.1099/0022-1317-57-2-307Google Scholar: Lookup
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  • Journal Article

Summary

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The research article examines the genome of an Australian strain of equine herpesvirus type 1, particularly in terms of its arrangement of cleavage sites for restriction enzymes EcoRI, BglII and BamHI. It employs a combination of techniques to establish a physical map of this genome, revealing two unique regions—one short and commonly found in either orientation in the DNA population, and one long, existing only in a single orientation.

Genome Analysis of Equine Herpesvirus Type 1

In this study, the researchers have focused on an isolated variant of equine herpesvirus type 1, colloquially known as equine abortion virus, from Australia. Their analysis primarily involved:

  • Assessing the genetic makeup or genome of this virus.
  • Studying the arrangement of sites on the genome where restriction endonucleases (enzymes that cut DNA at specific locations) can operate.
  • Formulating a physical map of this genome, which provides a visual representation of the virus’s genetic constitution and arrangement.

Use of Restriction Endonucleases

The researchers used three specific restriction endonucleases in their study:

  • EcoRI
  • BglII
  • BamHI

These enzymes are known for their ability to cleave DNA at particular sequences, making them instrumental in dissecting the genome of the herpesvirus.

Identification of Terminal Fragments and Linkage

The team used exonuclease treatments to identify terminal fragments. Exonucleases are enzymes that degrade DNA by removing nucleotides from the ends. This helped to identify the ends of the herpesvirus genome. They also deduced the linkage of various fragments or parts of the genome. This was achieved by a combination of:

  • Single- and double-digest experiments which help determine where the restriction sites are on the DNA (these are the places where the restriction enzymes mentioned earlier cut the DNA into pieces).
  • Cross-blot hybridizations – a technique used to confirm or study the relationship and similarities between two sequences of DNA.

Findings: Unique Regions in the Genome

The genome of equine herpesvirus was found to have a molecular weight of 100 x 106. The genome analysis revealed two unique segments:

  • A short unique region bounded by repetitive sequences. This region was present in both orientations, forward and reverse, approximately equally in the virus’s DNA population.
  • A long unique region that was found to exist only in a single orientation.

This orientation aspect in the herpesvirus genome is critical in understanding the functionality and life cycle of the virus, and has potential implications for disease control strategies, virus evolution, and the development of vaccines.

Cite This Article

APA
Whalley JM, Robertson GR, Davison AJ. (1981). Analysis of the genome of equine herpesvirus type 1: arrangement of cleavage sites for restriction endonucleases EcoRI, BglII and BamHI. J Gen Virol, 57(Pt 2), 307-323. https://doi.org/10.1099/0022-1317-57-2-307

Publication

ISSN: 0022-1317
NlmUniqueID: 0077340
Country: England
Language: English
Volume: 57
Issue: Pt 2
Pages: 307-323

Researcher Affiliations

Whalley, J M
    Robertson, G R
      Davison, A J

        MeSH Terms

        • Bacterial Proteins
        • DNA Restriction Enzymes
        • DNA, Viral / analysis
        • Deoxyribonuclease BamHI
        • Deoxyribonuclease EcoRI
        • Deoxyribonucleases, Type II Site-Specific
        • Genes, Viral
        • Herpesviridae / genetics
        • Herpesvirus 1, Equid / genetics
        • Molecular Weight
        • Nucleic Acid Hybridization
        • Repetitive Sequences, Nucleic Acid

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