Generation of a transposon insertion mutant library for bovine herpesvirus 4 cloned as a bacterial artificial chromosome by in vitro MuA based DNA transposition system.

Donofrio, Gaetano; Martignani, Eugenio; Sartori, Chiara; Vanderplasschen, Alain; Cavirani, Sandro; Flammini, Cesidio Filippo; Gillet, Laurent

doi:10.1016/j.jviromet.2006.11.028

Article (Scientific journals)

Generation of a transposon insertion mutant library for bovine herpesvirus 4 cloned as a bacterial artificial chromosome by in vitro MuA based DNA transposition system.

Donofrio, Gaetano; Martignani, Eugenio; Sartori, Chiara et al.

2007 • In Journal of Virological Methods, 141 (1), p. 63-70

Peer Reviewed verified by ORBi

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https://hdl.handle.net/2268/29080

DOI
10.1016/j.jviromet.2006.11.028

PubMed
17182112

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Keywords :

Animals; Catalysis; Cattle; Cell Line; Chromosomes, Artificial, Bacterial; Cloning, Molecular; DNA Transposable Elements; DNA, Viral; Electroporation; Escherichia coli/genetics; Gene Library; Herpesvirus 4, Bovine/genetics; Models, Genetic; Mutagenesis, Insertional; Transposases/metabolism

Abstract :

[en] Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus with no clear disease association. Although the BoHV-4 genome has been sequenced, the function of the majority of putative genes is elusive. Several features make BoHV-4 attractive as a backbone for use as a viral expression vector and/or as a model to study gamma herpesvirus biology and determining which genes are essential for its replication is a very important task. Starting from BoHV-4 genome cloned as infectious bacterial artificial chromosome (BAC-BoHV-4) in Escherichia coli. A random insertion mutant library for BoHV4 was generated by the use of MuA transposase-catalyzed in vitro transposition reaction. Viral mutant transfection and direct sequencing allow the rapid determination of which BoHV-4 genes are essential for viral growth in a permissive eukaryotic cell line. BoHV-4 functional analysis information is fundamental when the BoHV-4 genome is modified for vector purposes.

Disciplines :

Biochemistry, biophysics & molecular biology
Microbiology
Biotechnology

Author, co-author :

Donofrio, Gaetano

Martignani, Eugenio

Sartori, Chiara

Vanderplasschen, Alain ; Université de Liège - ULiège > Immunologie et vaccinologie

Cavirani, Sandro

Flammini, Cesidio Filippo

Gillet, Laurent ; Université de Liège - ULiège > Immunologie et vaccinologie

Language :

English

Title :

Generation of a transposon insertion mutant library for bovine herpesvirus 4 cloned as a bacterial artificial chromosome by in vitro MuA based DNA transposition system.

Publication date :

2007

Journal title :

Journal of Virological Methods

ISSN :

0166-0934

eISSN :

1879-0984

Publisher :

Elsevier Science, Amsterdam, Netherlands

Volume :

141

Issue :

Pages :

63-70

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 22 November 2010

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