Discrete Subgenomic DNA Fragments in Incomplete Particles of Adenovirus Type 2

Abstract

SUMMARYDiscrete subgenomic DNA fragments were found in three out of thirty-two preparations of adenovirus type 2 incomplete particles grown in human Hep-2 cells and examined over the course of 1 year. One preparation contained three fragments corresponding to 5, 14 and 19 % of the genome, another contained a 37 % fragment and the third a 40% fragment. Each fragment hybridized exclusively to the left end of the genome. Digestion of the nick-translated 37% fragment with HindIII confirmed that it contained the 1eft 37.3% of the genome. Synthesis of these fragments was not dependent on high input multiplicity of infection. Comparable fragments were not found in unpackaged DNA from the corresponding infected cells. This is consistent with the hypothesis that such fragments are generated during virus assembly or, alternatively, may reflect the very small proportion of these fragments relative to the pool of unpackaged DNA within the cells. The possibility that they are generated by errors in DNA replication is discussed.

Group C adenovirus DNA sequences in human lymphoid cells

“…High-molecular-weight DNA was extracted from the nuclei of PBL from 10 ml of fresh heparinized blood, digested to completion with Sall or BamHI and electrophoresed in a 1% agarose slab gel. The DNA was blotted onto nitrocellulose and hybridized to a 32P-labeled Ad2 probe (specific activity, 4 x 108 cpm/lg of DNA) as described previously (5). The probe consisted of an equimolar mixture of pBR322 plasmids containing the 10 Ad2 HindIII fragments.…”