Award details

Manipulation of the African swine fever virus genome

Reference	BBS/E/I/00001040
Principal Investigator / Supervisor	Dr Linda Dixon
Co-Investigators / Co-Supervisors
Institution	The Pirbright Institute
Department	The Pirbright Institute Department
Funding type	Research
Value (£)	1,403,846
Status	Completed
Type	Institute Project
Start date	01/04/2003
End date	30/06/2009
Duration	75 months

Abstract

Existing methods to manipulate the African swine fever virus genome rely on assay of reporter genes to identify recombinant viruses. These methods are time-consuming and inefficient as they depend on identification of recombinant viruses in a large background of wild-type viruses. This project will investigate approaches to improve the methods for constructing mutants ASFV viruses. One approach will be to introduce by homologous recombination a dominant selectable marker, the puromycin resistance gene, linked to the green fluorescent protein gene and cloned under control of an ASFV promoter (early and late promoters will be tested). Recombinant viruses will be selected by growing infected cells in the presence of puromycin and cells infected with recombinant viruses identified by expression of green fluorescent protein. Initial experiments will focus on constructing mutant viruses with the CD2v gene deleted or altered. We will also investigate the potential of using siRNAs to reduce expression of virus and cellular proteins. This approach could be useful to study the function of virus genes, which are essential for replication as well as to study the function of host proteins during virus replication. Virus genes encoding proteins against which we have antibodies will be targeted as well as host proteins which we have shown to bind to virus-encoded proteins. These include the SH3P7 protein which binds to the Cd2v protein and the ? NAC protein which binds the virus j4R protein. Functional effects of these mutations will also be studied.

Summary

unavailable

Committee	Closed Committee - Animal Sciences (AS)
Research Topics	Animal Health, Microbiology
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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