Construction of a gene inactivation library for Bovine herpesvirus 1 using infectious clone technology.
The application of infectious clone technology to herpesvirus biology has revolutionized the study of these viruses. Previously the ability to manipulate these large DNA viruses was limited to methods dependent on homologous recombination in mammalian cells. However, the construction of herpesvirus infectious clones using bacterial artificial chromosome vectors has permitted the application of powerful bacterial genetics for the manipulation of these viruses. A method is described for the construction and characterization of a gene inactivation library of Bovine herpesvirus 1 using an infectious clone. The method utilizes transposon-mediated gene inactivation, which permits gene inactivation without any prior knowledge of the viral genomic sequence. Furthermore, as the genetic manipulation is performed in bacteria the inactivation of those viral genes that are essential for viral replication is also possible. The method described here can be readily applied to any herpesvirus clone and provides the tools for precise characterization of all the genes contained within a herpesvirus genome.
Mahony
Timothy J
TJ
Department of Primary Industries, Agency for Food and Fibre Sciences, St. Lucia, Queensland, Australia.
McCarthy
Fiona M
FM
Gravel
Jennifer L
JL
Young
Peter L
PL
D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements; D015723:Gene Library; D020868:Gene Silencing; D005822:Genetic Vectors; D007242:Herpesvirus 1, Bovine / Q000235:genetics*; D010957:Plasmids
D004251:DNA Transposable Elements