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Restriction enzyme analysis of human cytomegalovirus using DNA extracted from infected cells

✍ Scribed by Lena Grillner; Ingrid Blomberg

Publisher
John Wiley and Sons
Year
1984
Tongue
English
Weight
645 KB
Volume
14
Category
Article
ISSN
0146-6615

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✦ Synopsis

Cytomegalovirus (CMV) isolates were investigated by restriction enzyme analysis. A method for extraction of cellular CMV DNA is described which uses CMV DNA released into the Hirt supernatant after precipitation of cellular DNA. CMV DNA is further purified by ultracentrifugation in neutral sucrose gradient. Fifteen strains from unrelated patients all had unique cleavage patterns with three restriction enzymes, although many of the fragments comigrated. Strains from related patients mostly showed identical patterns. The CMV genome seems to be stable since the pattern did not change in five isolates obtained from the same patient during a 5-year period.

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Numerous biological factors have an effect upon the probability of the presence of a theoretical restriction site at a given point, (1/4)n, in a DNA molecule, where n represents the number of base pairs composing the site. Indeed, it is a fact that the ratio (formula; see text) does not usually equa