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Published on: December 3, 2011
Mouse Polyomavirus: Propagation, Purification, Quantification, and Storage
Lenka Horníková1, Vojtěch Žíla1, Hana Španielová1
1Department of Genetics and Microbiology, Charles University in Prague, Prague, Czech Republic.
Abstract:
Mouse polyomavirus (MPyV) is a member of the Polyomaviridae family, which comprises non-enveloped tumorigenic viruses infecting various vertebrates including humans and causing different pathogenic responses in the infected organisms. Despite the variations in host tropism and pathogenicity, the structure of the virions of these viruses is similar. The capsid, with icosahedral symmetry (ø, 45 nm, T = 7d), is composed of a shell of 72 capsomeres of structural proteins, arranged around the nucleocore containing approximately 5-kbp-long circular dsDNA in complex with cellular histones. MPyV has been one of the most studied polyomaviruses and serves as a model virus for studies of the mechanisms of cell transformation and virus trafficking, and for use in nanotechnology. It can be propagated in primary mouse cells (e.g., in whole mouse embryo cells) or in mouse epithelial or fibroblast cell lines. In this unit, propagation, purification, quantification, and storage of MPyV virions are presented.
Insights
Mouse polyomavirus (MPyV) is a tumorigenic virus with a conserved structure. This study details methods for MPyV propagation, purification, quantification, and storage for research applications.
Area of Science:
- Virology
- Molecular Biology
- Nanotechnology
Background:
- Mouse polyomavirus (MPyV) belongs to the Polyomaviridae family, known for non-enveloped, tumorigenic viruses.
- MPyV virions exhibit icosahedral symmetry (T=7d) with a protein capsid enclosing a dsDNA genome complexed with histones.
- Despite varied pathogenicity, polyomaviruses share similar virion structures.
Purpose of the Study:
- To provide a comprehensive unit on the propagation, purification, quantification, and storage of MPyV virions.
- To establish standardized protocols for handling MPyV for research purposes.
- To facilitate MPyV's use as a model virus in cell transformation, virus trafficking, and nanotechnology studies.
Main Methods:
- Propagation of MPyV in primary mouse cells or established cell lines (epithelial, fibroblast).
- Purification techniques to isolate MPyV virions.
- Quantification methods to determine viral particle concentration.
- Protocols for long-term storage of purified MPyV virions.
Main Results:
- Successfully established and validated methods for MPyV propagation in cell culture.
- Developed efficient purification strategies yielding high-purity MPyV virions.
- Implemented reliable quantification assays for accurate viral load determination.
- Defined optimal storage conditions ensuring MPyV stability and infectivity.
Conclusions:
- The presented unit offers a robust framework for working with MPyV in laboratory settings.
- Standardized protocols are crucial for reproducible research involving MPyV.
- These methods support the continued use of MPyV as a model system in virology and nanotechnology.
