Abstract
KB cells coinfected with adenovirus and adeno-associated virus (AAV) yielded two kinds of infectious AAV particles that banded in CsCl at densities of 1.45 and 1.41 g/cm(2), respectively. The 1.45 band was found to be composed of a heterogeneous group of viral particles that could be subfractionated by velocity sedimentation. The main component from this band had a smaller S value (109) than the main component from the 1.41 band (111S), although both had the same DNA/protein ratio and the same density in metrizamide gradients. Continuous-label experiments showed that early after infection, both components (1.45 and 1.41) were generated in the same amounts, but this was followed by a relative increase in the proportion of the 1.41 component over the 1.45 particles. Pulse-chase analysis failed to demonstrate a precursor-product relationship between these two bands. The slower-sedimenting components from the 1.45 band were unstable in CsCl and were present in a greater proportion early after infection. These particles contained DNA that was enriched for the terminal sequences of the AAV genomes and was accessible to digestion with micrococcal nuclease.
MeSH Terms
Base Sequence
Cell Line
Centrifugation, Density Gradient
DNA Restriction Enzymes/pharmacology
DNA, Viral/analysis
Dependovirus/analysis,growth & development
Humans
Spectrophotometry, Ultraviolet
Virion/analysis
Chemicals
DNA, Viral
DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
de la Maza L M
Carter B J
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