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Herpesvirus genome mapping: a rapid generic approach.
1CRC for Conservation and Management of Marsupials, Queensland Department of Primary Industries, St Lucia, Australia. d.m.thomson@massey.ac.nz
Journal of Virological Methods
|October 3, 1999
Summary
A simplified protocol maps macropodid herpesvirus 1 genome using DNA sequencing and bioinformatics. This method accelerates viral genome analysis and gene ordering for herpesvirus research.
Area of Science:
- Virology
- Genomics
- Molecular Biology
Background:
- Macropodid herpesvirus 1 (MHV1) is an alphaherpesvirus affecting marsupials.
- Previous genome mapping protocols were complex and time-consuming.
Purpose of the Study:
- To describe a simplified and efficient protocol for mapping the MHV1 genome.
- To facilitate rapid gene ordering and analysis within large viral clones.
Main Methods:
- Viral DNA undergoes single restriction digestion and cloning into a plasmid vector.
- Fragment ends are sequenced, translated, and searched against peptide databases.
- Putative contiguous fragments are assembled using shared open reading frames and PCR confirmation.
- Unidentified fragments are rapidly subcloned and sequenced.
Main Results:
- The protocol successfully maps the MHV1 genome with simplified steps.
- Rapid subcloning aids in sequencing and ordering genes within large clones.
- The method leverages sequence homology and colinearity for map construction.
Conclusions:
- This protocol offers a significant improvement for alphaherpesvirus genome mapping.
- The approach is adaptable for gene ordering and analysis in related viruses.
- This research provides a valuable tool for understanding herpesvirus biology.