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altering the ad5 packaging domain affects the maturation of the ad particles改变ad5包装领域影响广告的成熟粒子
Altering the Ad5 Packaging Domain Affects the
Maturation of the Ad Particles
1¤ 1 2 1 2 1,3
Raul Alba , Dan Cots , Philomena Ostapchuk , Assumpcio Bosch , Patrick Hearing , Miguel Chillon *
`
1 Center of Animal Biotechnology and Gene Therapy (CBATEG), and Department of Biochemistry and Molecular Biology, Universitat Autonoma de Barcelona, Bellaterra,
Barcelona, Spain, 2 Department of Molecular Genetics and Microbiology, School of Medicine, Stony Brook University, Stony Brook, New York, United States of America,
`
3 Institut Catala de Recerca i Estudis Avanc¸ats (ICREA), Barcelona, Spain
Abstract
We have previously described a new family of mutant adenoviruses carrying different combinations of attB/attP sequences
from bacteriophage PhiC31 flanking the Ad5 packaging domain. These novel helper viruses have a significantly delayed viral
life cycle and a severe packaging impairment, regardless of the presence of PhiC31 recombinase. Their infectious viral titers
are significantly lower (100–1000 fold) than those of control adenovirus at 36 hours post-infection, but allow for efficient
packaging of helper-dependent adenovirus. In the present work, we have analyzed which steps of the adenovirus life cycle
are altered in attB-helper adenoviruses and investigated whether these viruses can provide the necessary viral proteins in
trans. The entry of attB-adenoviral genomes into the cell nucleus early at early timepoints post-infection was not impaired
and viral protein expression levels were found to be similar to those of control ade
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