Generation of E3-deleted canine adenoviruses expressing canine parvovirus capsid by homologous recombination in bacteria.
E3-deleted canine adenovirus type 1 (CAV-1) was generated by homologous recombination in bacterial cells, using an antibiotic resistance marker to facilitate the recovery of recombinants. This marker was flanked by unique restriction endonuclease sites, which allowed its subsequent removal and the insertion of cassettes expressing the canine parvovirus capsid at the E3 locus. Infectious virus was recovered following transfection of canine cells and capsid expression was observed by RT-PCR from one of the virus constructs. A second construct, containing a different promoter, showed delayed growth and genome instability which, based on the size difference between these inserts, suggests a maximum packaging size of 106 to 109% wild-type genome size for CAV-1.
Morrison
Mark D
MD
Department of Veterinary Pathology, University of Glasgow Veterinary School, Garscube Estate, Glasgow G61 1QH, United Kingdom. m.morrison@vet.gla.ac.uk
Reid
Dorothy
D
Onions
David
D
Spibey
Norman
N
Nicolson
Lesley
L
D017503:Adenovirus E3 Proteins / Q000235:genetics*; D006516:Adenoviruses, Canine / Q000235:genetics*; D000818:Animals; D001419:Bacteria / Q000235:genetics; D002213:Capsid / Q000235:genetics*; D002460:Cell Line; D004285:Dogs; D017993:Parvovirus, Canine / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic
D017503:Adenovirus E3 Proteins