Evaluation of the Deletion of the African Swine Fever Virus Gene <i>O174L</i> from the Genome of the Georgia Isolate
2023
Elizabeth Ramirez-Medina | Lauro Velazquez-Salinas | Ayushi Rai | Nallely Espinoza | Alyssa Valladares | Ediane Silva | Leeanna Burton | Edward Spinard | Amanda Meyers | Guillermo Risatti | Sten Calvelage | Sandra Blome | Douglas P. Gladue | Manuel V. Borca
African swine fever virus (ASFV) is a structurally complex, double-stranded DNA virus, which causes African swine fever (ASF), a contagious disease affecting swine. ASF is currently affecting pork production in a large geographical region, including Eurasia and the Caribbean. ASFV has a large genome, which harbors more than 160 genes, but most of these genes’ functions have not been experimentally characterized. One of these genes is the <i>O174L</i> gene which has been experimentally shown to function as a small DNA polymerase. Here, we demonstrate that the deletion of the <i>O174L</i> gene from the genome of the virulent strain ASFV Georgia2010 (ASFV-G) does not significantly affect virus replication in vitro or in vivo. A recombinant virus, having deleted the <i>O174L</i> gene, ASFV-G-∆O174L, was developed to study the effect of the <i>O174L</i> protein in replication in swine macrophages cultures in vitro and disease production when inoculated in pigs. The results demonstrated that ASFV-G-∆O174L has similar replication kinetics to parental ASFV-G in swine macrophage cultures. In addition, animals intramuscularly inoculated with 10<sup>2</sup> HAD<sub>50</sub> of ASFV-G-∆O174L presented a clinical form of the disease that is indistinguishable from that induced by the parental virulent strain ASFV-G. All animals developed a lethal disease, being euthanized around day 7 post-infection. Therefore, although O174L is a well-characterized DNA polymerase, its function is apparently not critical for the process of virus replication, both in vitro and in vivo, or for disease production in domestic pigs.
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