Current Topics in Microbiology and Immunology by Malcolm R. Macnaughton (auth.), W. Henle, P. H.

By Malcolm R. Macnaughton (auth.), W. Henle, P. H. Hofschneider, H. Koprowski, F. Melchers, R. Rott, H. G. Schweiger, P. K. Vogt (eds.)

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It is now recognized that the successful extraction and purification of intact, fulllength genomes from poxvirions not only requires careful attention to the conditions employed for releasing DNA from the virus particles and the subsequent manipulations involved in deproteinization of the DNA, but one must also be cognizant of the manipulations and conditions employed during the purification of the virus particles from which the DNA will be extracted (Holowczak 1976). Repeated freezing and thawing and(or) sonication of virus or cell extracts containing virus appear to cause or predispose the genome, while still encapsulated within virions, to become "nicked" or fragmented (Fig.

1977) it seems reasonable to propose that the core region, observed in the electron microscope to be present in nucleoids and Gu-subviral particles, may represent the central region of the ftgure eight-structure or spool into which the poxvirus genome is folded. Since the spool is twisted 1800 , proteins could be required to stabilize the structure at the point where it is twisted and these proteins in combination with DNA would form the densely staining central core region observed in the electron microscope.

For this to be effective, a collection of well-characterized stocks of vaccinia virus should be retained and be available to investigators. The American Type Culture Collection does maintain a few strains of vaccinia virus in their collection, but it would be necessary to fully characterize these strains, including restriction enzyme analysis of the DNA they contain, for the collection to be useful in this regard. General agreement would also be necessary as to what would constitute the restriction enzyme pattern "standard" for a particular strain of vaccinia virus or other poxviruses.

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