Canadian Forest Service Publications
Isolation of an Entomopoxvirus and Characterization of its DNA. 1976. Arif, B.M. Virology 69: 626 - 634.
Issued by: Great Lakes Forestry Centre
Catalog ID: 28896
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An entomopoxvirus from Choristoneura biennis was propagated in the eastern spruce budworm, Choristoneura fumiferana. The virus, released from its inclusion bodies by alkaline treatment, sedimented as a homogeneous band by rate zonal centrifugation through sucrose gradients. Electron microscopic examination showed that the particles possessed the mulberrylike structure that is typical of intact entomopox virions. The virus also produced a single band by isopycnic centrifugation in CsCl at a density of 1.31 g/ml. The genome, extracted with sodium lauryl sarcosinate and Pronase, sedimented as a single band through sucrose gradients, and its MW was calculated to be 132-142 × 106 when compared to marker DNA from coliphage T2. Its sedimentation coefficient was 59S. The G + C content of the viral genome was investigated by thermal denaturation and buoyant density analysis in CsCl. The DNA melted at 61.6° and had a density of 1.6849 g/ml. These values are equivalent to a G + C content of 26.6% and 25.4%, respectively, which are considerably lower than those reported for the genomes of vertebrate poxviruses (G + C, 32.5–39%). The difference in G + C content between vertebrate and invertebrate poxviruses was confirmed by characterizing the genome of a second entomopoxvirus with respect to its tm and buoyant density. The double-stranded nature of the DNA from invertebrate poxviruses was established from melting profiles and reaction with formaldehyde.