ORBi: Costes Bérénice - Cloning of the koi herpesvirus genome as an infectious bacterial artificial chromosome demonstrates that disruption of the thymidine kinase locus induces partial attenuation in Cyprinus carpio koi.

Reference : Cloning of the koi herpesvirus genome as an infectious bacterial artificial chromosome d...


	Document type :	Scientific journals : Article
	Discipline(s) :	Life sciences : Veterinary medicine & animal health
	To cite this reference:	http://hdl.handle.net/2268/18037

Title :	Cloning of the koi herpesvirus genome as an infectious bacterial artificial chromosome demonstrates that disruption of the thymidine kinase locus induces partial attenuation in Cyprinus carpio koi.
Language :	English
Author, co-author :	Costes, Bérénice [Université de Liège - ULg > > Immunologie et vaccinologie >]
	Fournier, Guillaume [Université de Liège - ULg > > Immunologie et vaccinologie >]
	Michel, Benjamin [Université de Liège - ULg > > Immunologie et vaccinologie >]
	Delforge, Cédric [Université de Liège - ULg > > Immunologie et vaccinologie >]
	Stalin Raj, V. [> >]
	Dewals, Benjamin G [Université de Liège - ULg > > Immunologie et vaccinologie >]
	Gillet, Laurent [Université de Liège - ULg > > Immunologie et vaccinologie >]
	Drion, Pierre [Université de Liège - ULg > Services généraux (Faculté de médecine vétérinaire) > Méth. expér. des anim. de labo et éthique en expér. animale >]
	Body, A. [> >]
	Schynts, F. [> >]
	Lieffrig, F. [> >]
	Vanderplasschen, Alain [Université de Liège - ULg > > Immunologie et vaccinologie >]
Publication date :	2008
Journal title :	Journal of Virology
Publisher :	American Society for Microbiology (ASM)
Volume :	82
Issue/season :	10
Pages :	4955-4964
Audience :	International
ISSN :	0022-538X
e-ISSN :	1098-5514
City :	Washington
Country :	DC
Abstract :	[en] Koi herpesvirus (KHV) is the causative agent of a lethal disease in koi and common carp. In the present study, we describe the cloning of the KHV genome as a stable and infectious bacterial artificial chromosome (BAC) clone that can be used to produce KHV recombinant strains. This goal was achieved by the insertion of a loxP-flanked BAC cassette into the thymidine kinase (TK) locus. This insertion led to a BAC plasmid that was stably maintained in bacteria and was able to regenerate virions when permissive cells were transfected with the plasmid. Reconstituted virions free of the BAC cassette but carrying a disrupted TK locus (the FL BAC-excised strain) were produced by the transfection of Cre recombinase-expressing cells with the BAC. Similarly, virions with a wild-type revertant TK sequence (the FL BAC revertant strain) were produced by the cotransfection of cells with the BAC and a DNA fragment encoding the wild-type TK sequence. Reconstituted recombinant viruses were compared to the wild-type parental virus in vitro and in vivo. The FL BAC revertant strain and the FL BAC-excised strain replicated comparably to the parental FL strain. The FL BAC revertant strain induced KHV infection in koi carp that was indistinguishable from that induced by the parental strain, while the FL BAC-excised strain exhibited a partially attenuated phenotype. Finally, the usefulness of the KHV BAC for recombination studies was demonstrated by the production of an ORF16-deleted strain by using prokaryotic recombination technology. The availability of the KHV BAC is an important advance that will allow the study of viral genes involved in KHV pathogenesis, as well as the production of attenuated recombinant candidate vaccines.
Target :	Researchers ; Professionals
Permalink :	http://hdl.handle.net/2268/18037
	also: http://hdl.handle.net/2268/29061
DOI :	10.1128/JVI.00211-08
Other URL :	http://jvi.asm.org/cgi/reprint/82/10/4955

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