Establishment of a Simple and Efficient Reverse Genetics System for Canine Adenoviruses Using Bacterial Artificial Chromosomes.
Adenoviridae Infections
/ veterinary
Adenoviruses, Canine
/ genetics
Animals
Chromosomes, Artificial, Bacterial
/ genetics
Cloning, Molecular
Dogs
Genome, Viral
/ genetics
Hepatitis, Infectious Canine
/ virology
High-Throughput Nucleotide Sequencing
Madin Darby Canine Kidney Cells
/ virology
Reverse Genetics
/ methods
adenovirus
bacterial artificial chromosome
canine
reverse genetics
vector
Journal
Viruses
ISSN: 1999-4915
Titre abrégé: Viruses
Pays: Switzerland
ID NLM: 101509722
Informations de publication
Date de publication:
16 07 2020
16 07 2020
Historique:
received:
01
07
2020
revised:
14
07
2020
accepted:
14
07
2020
entrez:
26
7
2020
pubmed:
28
7
2020
medline:
23
2
2021
Statut:
epublish
Résumé
Canine adenoviruses (CAdVs) are divided into pathotypes CAdV1 and CAdV2, which cause infectious hepatitis and laryngotracheitis in canid animals, respectively. They can be the backbones of viral vectors that could be applied in recombinant vaccines or for gene transfer in dogs and in serologically naïve humans. Although conventional plasmid-based reverse genetics systems can be used to construct CAdV vectors, their large genome size creates technical difficulties in gene cloning and manipulation. In this study, we established an improved reverse genetics system for CAdVs using bacterial artificial chromosomes (BACs), in which genetic modifications can be efficiently and simply made through BAC recombineering. To validate the utility of this system, we used it to generate CAdV2 with the early region 1 gene deleted. This mutant was robustly generated and attenuated in cell culture. The results suggest that our established BAC-based reverse genetics system for CAdVs would be a useful and powerful tool for basic and advanced practical studies with these viruses.
Identifiants
pubmed: 32708703
pii: v12070767
doi: 10.3390/v12070767
pmc: PMC7412426
pii:
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
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