title: A novel bromodeoxyuridine-resistant wild boar lung cell line facilitates generation of African swine fever virus recombinants.
abstract: Manipulation of African swine fever virus (ASFV) genomes, in particular those from field strains, is still a challenge. We have shown recently that generation of a green-fluorescent-protein-expressing, thymidine-kinase-negative (TK-) mutant of the low-pathogenic African swine fever virus field strain NHV was supported by a TK- Vero cell line. Since NHV, like other ASFV field strains, does not replicate well in Vero cells, a bromodeoxyuridine (BrdU)- resistant cell line derived from wild boar lung (WSL) cells, named WSL-Bu, was selected. WSL cells were used because they are suitable for productive replication of NHV and other ASFV field strains. Here, we show that WSL-Bu cells enable positive selection of both TK- and TK+ ASFV recombinants, which allows for novel strategies for construction of ASFV mutants. We further demonstrate for a low-pathogenic ASFV strain that TK expression is required for infectious replication in macrophages infected at low multiplicity and that vaccinia TK fully complements ASFV TK in this respect.
authors.affiliation: Institut für molekulare Virologie und Zellbiologie, Friedrich-Loeffler-Institut, 17493, Greifswald-Insel Riems, Germany, guenther.keil@fli.bund.de.
authors.forename: Günther M
authors.initials: GM
authors.lastname: Keil
authors.forename: Katrin
authors.initials: K
authors.lastname: Giesow
authors.forename: Raquel
authors.initials: R
authors.lastname: Portugal
mesh_terms: D000358:African Swine Fever Virus / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification / Q000502:physiology; D000818:Animals; D002460:Cell Line; D008168:Lung; D011995:Recombination, Genetic; D012641:Selection, Genetic; D034421:Sus scrofa; D013937:Thymidine Kinase / Q000378:metabolism; D014776:Virus Cultivation / Q000379:methods; D014779:Virus Replication
chemical_list: D013937:Thymidine Kinase