Article (Scientific journals)
Development of bovine herpesvirus 4 as an expression vector using bacterial artificial chromosome cloning.
Gillet, Laurent; Daix, Virginie; Donofrio, G. et al.
2005In Journal of General Virology, 86 (Pt 4), p. 907-17
Peer Reviewed verified by ORBi
 

Files


Full Text
Development of bovine herpesvirus 4 as an expression vector using bacterial artificial chromosome cloning -- Gillet et al_ 86 (4) 907.pdf
Author postprint (818.91 kB)
Request a copy

All documents in ORBi are protected by a user license.

Send to



Details



Keywords :
Animals; Cattle; Chromosomes, Artificial, Bacterial; Cloning, Molecular; Complement Inactivator Proteins/genetics/metabolism; Escherichia coli/genetics/metabolism; Genetic Vectors; Herpesvirus 4, Bovine/genetics/metabolism/physiology; Ixodes/immunology/metabolism; Recombination, Genetic; Virus Replication
Abstract :
[en] Several features make bovine herpesvirus 4 (BoHV-4) attractive as a backbone for use as a viral expression vector and/or as a model to study gammaherpesvirus biology. However, these developments have been impeded by the difficulty in manipulating its large genome using classical homologous recombination in eukaryotic cells. In the present study, the feasibility of exploiting bacterial artificial chromosome (BAC) cloning and prokaryotic recombination technology for production of BoHV-4 recombinants was explored. Firstly, the BoHV-4 genome was BAC cloned using two potential insertion sites. Both sites of insertion gave rise to BoHV-4 BAC clones stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. Reconstituted virus replicated comparably to wild-type parental virus and the loxP-flanked BAC cassette was excised by growing them on permissive cells stably expressing Cre recombinase. Secondly, BoHV-4 recombinants expressing Ixodes ricinus anti-complement protein I or II (IRAC I/II) were produced using a two-step mutagenesis procedure in Escherichia coli. Both recombinants induced expression of high levels of functional IRAC molecules in the supernatant of infected cells. This study demonstrates that BAC cloning and prokaryotic recombination technology are powerful tools for the development of BoHV-4 as an expression vector and for further fundamental studies of this gammaherpesvirus.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Gillet, Laurent  ;  Université de Liège - ULiège > Immunologie et vaccinologie
Daix, Virginie 
Donofrio, G.
Wagner, M.
Koszinowski, U. H.
China, B.
Ackermann, M.
Markine-Goriaynoff, Nicolas 
Vanderplasschen, Alain ;  Université de Liège - ULiège > Immunologie et vaccinologie
Language :
English
Title :
Development of bovine herpesvirus 4 as an expression vector using bacterial artificial chromosome cloning.
Publication date :
2005
Journal title :
Journal of General Virology
ISSN :
0022-1317
eISSN :
1465-2099
Publisher :
Society for General Microbiology, London, United Kingdom
Volume :
86
Issue :
Pt 4
Pages :
907-17
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 24 November 2009

Statistics


Number of views
54 (7 by ULiège)
Number of downloads
5 (5 by ULiège)

Scopus citations®
 
44
Scopus citations®
without self-citations
16
OpenCitations
 
36

Bibliography


Similar publications



Contact ORBi