title: Approaches to the identification of non-essential genes of African swine fever virus.
abstract: It is poorly understood why vaccines could not be developed for the control and prevention of African swine fever (ASF) virus infection. The aim of our study was to identify genes non-essential for ASF virus replication because there were indications that certain viral gene products, which apparently are non-essential for viral replication, conferred protection from death due to ASF. A cosmid library representing the genome of ASF virus strain France 64 was established and characterized. Then, in order to inactivate viral genes by insertion, the beta-galactosidase (beta-gal) gene was introduced either randomly or at specific locations of selected cloned DNA fragments. These constructions were transfected into cells which had been previously infected with a cell-culture-adapted viral strain in order to allow the generation of recombinant progeny virus. Viable recombinant progeny was identified by at least one of the following means: (1) expression of beta-gal; (2) detection of beta-gal specific DNA by plaque hybridization, and (3) absence of a functional product of the inactivated gene. Presently, we are characterizing a recombinant virus with an insertionally inactivated thymidine kinase gene.
authors: Institute for Viral Diseases, Basel, Switzerland.
authors: C
authors: C
authors: Mettraux
authors: M
authors: M
authors: Ackermann
authors: J D
authors: JD
authors: Tratschin
authors: U
authors: U
authors: Kihm
mesh_terms: D000358:African Swine Fever Virus / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003360:Cosmids; D004279:DNA, Viral / Q000032:analysis*; D015967:Gene Expression Regulation, Viral; D015723:Gene Library; D005814:Genes, Viral; D016254:Mutagenesis, Insertional; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D015183:Restriction Mapping; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection; D014779:Virus Replication / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D004279:DNA, Viral; D013937:Thymidine Kinase; D001616:beta-Galactosidase