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Restriction endonuclease mapping of the fowlpox virus genome
1CSIRO Australian Animal Health Laboratory, Geelong, Victoria.
Abstract:
The genome of fowlpox virus (Webster's mild vaccine strain) is composed of a single, double-stranded DNA molecule with covalently linked terminal hairpins and approximately 300 kb in length. Sites for cleavage by restriction endonucleases Pstl, Sal/l, Smal, and Notl have been identified with partial maps for EcoRI and BamHI. Differences in Pstl restriction fragment profiles for two separately prepared viruses (FPV-M and FPV-M3), both derived from the vaccine strain, indicate the presence of a nonessential region and potential insertion site for foreign DNA, toward one end of the viral genome. The size of the fowlpox virus genome, which is more than 100 kb larger than the orthopoxvirus vaccinia, indicates that the avipox viruses have the potential to code for more proteins than other groups of poxviruses.
Insights
The fowlpox virus genome is large, offering potential for more proteins. Differences in its DNA suggest a nonessential region suitable for foreign DNA insertion.
Area of Science:
- Virology
- Molecular Biology
- Genomics
Background:
- Fowlpox virus (FPV) is an avipoxvirus with a large double-stranded DNA genome.
- Understanding FPV genome structure is crucial for vaccine development and genetic manipulation.
Purpose of the Study:
- To characterize the genome of the FPV Webster's mild vaccine strain.
- To identify potential sites for foreign DNA insertion within the FPV genome.
Main Methods:
- Restriction endonuclease digestion (PstI, SalI, SmaI, NotI, EcoRI, BamHI) was used to map the FPV genome.
- Analysis of restriction fragment profiles differentiated viral strains.
Main Results:
- The FPV genome is approximately 300 kb, larger than vaccinia virus.
- Restriction sites for several endonucleases were identified, with partial maps for EcoRI and BamHI.
- Differences in PstI profiles between FPV-M and FPV-M3 strains suggest a nonessential genomic region.
Conclusions:
- The FPV genome's size indicates a greater coding capacity compared to orthopoxviruses.
- A nonessential region identified in the FPV genome presents a potential site for foreign DNA integration.