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Molecular cloning, physical mapping and cross-hybridization of the murine adenovirus type 1 and type 2 genomes
C Jacques1, L Cousineau, B D'Amours
1Centre de Recherche en Virologie, Institut Armand-Frappier, Laval-des-Rapides, Québec, Canada.
Abstract:
Murine adenovirus (MAd) type 1 strain FL and type 2 strain K87 genomes were cloned into plasmid pAT153 as HindIII restriction fragments. The MAd-1 and MAd-2 DNA genomes, 30.10 kb and 34.71 kb in length respectively, were mapped using BglII, ClaI, EcoRI, HindIII and SphI restriction endonuclease cleavage sites. In view of the large differences found between the MAd-1 and MAd-2 genomes in terms of the number and location of restriction sites, cross-hybridization experiments were performed. Homologous DNA sequences were located on the MAd-1 and MAd-2 physical maps. Both viruses are also genetically related to human adenovirus type 2 (HAd-2). Nucleotide sequences shared by HAd-2 and the MAds code for structural proteins, which may explain the antigenic similarities between these viruses from different origins. Our results confirm the existence of two distinct adenovirus species in the mouse.
Insights
Murine adenoviruses (MAd) type 1 and 2 genomes were analyzed, revealing distinct genetic profiles and homologous DNA sequences. These findings confirm two separate mouse adenovirus species with genetic links to human adenovirus type 2.
Area of Science:
- Virology
- Molecular Biology
- Genomics
Background:
- Murine adenoviruses (MAd) are known pathogens in mice.
- Understanding their genetic diversity is crucial for diagnostics and research.
- Previous studies suggested potential relationships between different MAd strains.
Purpose of the Study:
- To genetically characterize Murine Adenovirus type 1 (MAd-1) and Murine Adenovirus type 2 (MAd-2).
- To compare the restriction endonuclease cleavage sites and map the genomes of MAd-1 and MAd-2.
- To investigate the genetic relatedness between MAd-1, MAd-2, and human adenovirus type 2 (HAd-2).
Main Methods:
- Cloning of MAd-1 and MAd-2 genomes into plasmid pAT153.
- Restriction endonuclease digestion and analysis using BglII, ClaI, EcoRI, HindIII, and SphI.
- Cross-hybridization experiments to identify homologous DNA sequences.
- Physical mapping of viral genomes.
Main Results:
- The MAd-1 and MAd-2 genomes were mapped, showing significant differences in restriction site number and location (30.10 kb and 34.71 kb, respectively).
- Homologous DNA sequences were identified between MAd-1 and MAd-2.
- Genetic relatedness was established between MAd-1, MAd-2, and HAd-2, particularly in sequences coding for structural proteins.
Conclusions:
- The study confirms the existence of two distinct adenovirus species in mice: MAd-1 and MAd-2.
- Shared nucleotide sequences between MAd and HAd-2 likely explain observed antigenic similarities.
- These findings provide a foundation for further research into MAd pathogenesis and evolution.