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The control of African swine fever (ASF) has been hampered by the unavailability of vaccines. Successful experimental vaccines have been derived from naturally occurring, cell culture-adapted, or genetically modified live attenuated ASFV. Recombinant viruses harboring engineered deletions of specific virulence-associated genes induce solid protection against challenge with parental viruses. Deletion of the<jats:italic>9GL<\/jats:italic>(<jats:italic>B119L<\/jats:italic>) gene in the highly virulent ASFV isolates Malawi Lil-20\/1 (Mal) and Pretoriuskop\/96\/4 (\u03949GL viruses) resulted in complete protection when challenged with parental isolates. When similar deletions were created within the ASFV Georgia 2007 (ASFV-G) genome, attenuation was achieved but the protective and lethal doses were too similar. To enhance attenuation of ASFV-G, we deleted another gene,<jats:italic>UK<\/jats:italic>(<jats:italic>DP96R<\/jats:italic>), which was previously shown to be involved in attenuation of the ASFV E70 isolate. Here, we report the construction of a double-gene-deletion recombinant virus, ASFV-G-\u03949GL\/\u0394UK. When administered intramuscularly (i.m.) to swine, there was no induction of disease, even at high doses (10<jats:sup>6<\/jats:sup>HAD<jats:sub>50<\/jats:sub>). Importantly, animals infected with 10<jats:sup>4<\/jats:sup>50% hemadsorbing doses (HAD<jats:sub>50<\/jats:sub>) of ASFV-G-\u03949GL\/\u0394UK were protected as early as 14 days postinoculation when challenged with ASFV-G. The presence of protection correlates with the appearance of serum anti-ASFV antibodies, but not with virus-specific circulating ASFV-specific gamma interferon (IFN-\u03b3)-producing cells. ASFV-G-\u03949GL\/\u0394UK is the first rationally designed experimental ASFV vaccine that protects against the highly virulent ASFV Georgia 2007 isolate as early as 2 weeks postvaccination.<\/jats:p><jats:p><jats:bold>IMPORTANCE<\/jats:bold>Currently, there is no commercially available vaccine against African swine fever. Outbreaks of the disease are devastating to the swine industry and are caused by circulating strains of African swine fever virus. Here, we report a putative vaccine derived from a currently circulating strain but containing two deletions in two separate areas of the virus, allowing increased safety. Using this genetically modified virus, we were able to vaccinate swine and protect them from developing ASF. We were able to achieve protection from disease as early as 2 weeks after vaccination, even when the pigs were exposed to a higher than normal concentration of ASFV.<\/jats:p>","DOI":"10.1128\/jvi.01760-16","type":"journal-article","created":{"date-parts":[[2016,10,27]],"date-time":"2016-10-27T02:49:38Z","timestamp":1477536578000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":181,"title":["Simultaneous Deletion of the<i>9GL<\/i>and<i>UK<\/i>Genes from the African Swine Fever Virus Georgia 2007 Isolate Offers Increased Safety and Protection against Homologous Challenge"],"prefix":"10.1128","volume":"91","author":[{"given":"Vivian","family":"O'Donnell","sequence":"first","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"},{"name":"Department of Pathobiology and Veterinary Science, CANR, University of Connecticut, Storrs, Connecticut, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillermo R.","family":"Risatti","sequence":"additional","affiliation":[{"name":"Department of Pathobiology and Veterinary Science, CANR, University of Connecticut, Storrs, Connecticut, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauren G.","family":"Holinka","sequence":"additional","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter W.","family":"Krug","sequence":"additional","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jolene","family":"Carlson","sequence":"additional","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"},{"name":"Biosecurity Research Institute and Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauro","family":"Velazquez-Salinas","sequence":"additional","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul A.","family":"Azzinaro","sequence":"additional","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas P.","family":"Gladue","sequence":"additional","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manuel V.","family":"Borca","sequence":"additional","affiliation":[{"name":"Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Greenport, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"43","article-title":"African swine fever virus","volume":"328","author":"Tulman ER","year":"2009","unstructured":"Tulman ER Delhon GA Ku BK Rock DL . 2009. 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