Article (Scientific journals)
Cloning of the genome of Alcelaphine herpesvirus 1 as an infectious and pathogenic bacterial artificial chromosome.
Dewals, Benjamin G; Boudry, Christel; Gillet, Laurent et al.
2006In Journal of General Virology, 87 (Pt 3), p. 509-17
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Keywords :
Animals; Cattle; Cell Line; Chromosomes, Artificial, Bacterial/genetics; Cloning, Molecular; Escherichia coli/genetics; Gammaherpesvirinae/genetics; Genetic Vectors; Genome, Viral; Malignant Catarrh/virology; Rabbits; Transformation, Bacterial; Virion/pathogenicity/physiology; Virulence; Virus Replication
Abstract :
[en] Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order Artiodactyla. The study of MCF pathogenesis has been impeded by an inability to produce recombinant virus, mainly due to the fact that AlHV-1 becomes attenuated during passage in culture. In this study, these difficulties were overcome by cloning the entire AlHV-1 genome as a stable, infectious and pathogenic bacterial artificial chromosome (BAC). A modified loxP-flanked BAC cassette was inserted in one of the two large non-coding regions of the AlHV-1 genome. This insertion allowed the production of an AlHV-1 BAC clone stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. The loxP-flanked BAC cassette was excised from the genome of reconstituted virions by growing them in permissive cells stably expressing Cre recombinase. Importantly, BAC-derived AlHV-1 virions replicated comparably to the virulent (low-passage) AlHV-1 parental strain and induced MCF in rabbits that was indistinguishable from that of the virulent parental strain. The availability of the AlHV-1 BAC is an important advance for the study of MCF that will allow the identification of viral genes involved in MCF pathogenesis, as well as the production of attenuated recombinant candidate vaccines.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Dewals, Benjamin G  ;  Université de Liège - ULiège > Immunologie et vaccinologie
Boudry, Christel ;  Université de Liège - ULiège > Immunologie et vaccinologie
Gillet, Laurent  ;  Université de Liège - ULiège > Immunologie et vaccinologie
Markine-Goriaynoff, Nicolas 
De Leval, Laurence ;  Centre Hospitalier Universitaire de Liège - CHU > Anatomie pathologique
Haig, D. M.
Vanderplasschen, Alain ;  Université de Liège - ULiège > Immunologie et vaccinologie
Language :
English
Title :
Cloning of the genome of Alcelaphine herpesvirus 1 as an infectious and pathogenic bacterial artificial chromosome.
Publication date :
2006
Journal title :
Journal of General Virology
ISSN :
0022-1317
eISSN :
1465-2099
Publisher :
Society for General Microbiology, London, United Kingdom
Volume :
87
Issue :
Pt 3
Pages :
509-17
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 24 November 2009

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