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  • Mouse Polyomavirus: Propagation, Purification, Quantification, and Storage.

Mouse Polyomavirus: Propagation, Purification, Quantification, and Storage.

Current protocols in microbiology (2015-08-04)
Lenka Horníková, Vojtěch Žíla, Hana Španielová, Jitka Forstová
摘要

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.

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Sigma-Aldrich
牛血清白蛋白 来源于牛血清, cold ethanol fraction, pH 5.2, ≥96%
Sigma-Aldrich
明胶 来源于猪皮肤, gel strength 300, Type A
Sigma-Aldrich
抑肽酶 来源于牛肺, saline solution, 3-7 TIU/mg protein
Sigma-Aldrich
神经氨酸酶 来源于霍乱弧菌, Type III, buffered aqueous solution, 0.2 μm filtered, 1-5 units/mg protein (Lowry, using NAN-lactose)