Problems of VirologyProblems of Virology0507-40882411-2097Central Research Institute for Epidemiology910.36233/0507-4088-2019-64-4-193-200ASF virus replication features in the presence of recombinant proteins CD2v, pX69R and pE248RMazloumA.<p class="MsoNormal"><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Mazloum Ali - Ph.D.</span></p><p class="MsoNormal"><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Vladimir region, Vladimir city, Yuryevets microdistrict, 600901.</span></p>ali.mazloum6@gmail.comhttps://orcid.org/0000-0002-5982-8393ZhukovI. U.<p>Vladimir region, Vladimir city, Yuryevets microdistrict, 600901.</p>fake@neicon.ruhttps://orcid.org/0000-0002-3817-2129AronovaE. B.<p>Vladimir region, Vladimir city, Yuryevets microdistrict, 600901.</p>fake@neicon.ruhttps://orcid.org/0000-0002-2072-6701IgolkinA. S.<p>Vladimir region, Vladimir city, Yuryevets microdistrict, 600901.</p>fake@neicon.ruhttps://orcid.org/0000-0002-5438-8026VlasovaN. N.<p>Vladimir region, Vladimir city, Yuryevets microdistrict, 600901.</p>fake@neicon.ruhttps://orcid.org/0000-0001-8707-7710«ARRIAH» Federal State Budgetary Institution «Federal Center for Animal Health»200820196441932001712201917122019Copyright © 2019, Mazloum A., Zhukov I.U., Aronova E.B., Igolkin A.S., Vlasova N.N.2019<p class="MsoNormal"><strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Introduction</span></strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">. African swine fever (ASF), sever hemorrhagic disease of swine caused by a large DNA virus of the <em>Asfaviridae</em> family.</span></p><p class="MsoNormal"><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Since there are no effective and safe vaccines against ASF yet, it is urgent to study the functions of its proteins, which is applicable by analyzing the features of ASF virus replication in the presence of recombinant proteins in vitro.</span></p><p class="MsoNormal"><strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Purpose</span></strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">. To study the effect of ASFV recombinant proteins CD2v, pE248R and pX69R on the speed and level of reproduction of ASF virus <em>in vitro</em>. Thus, obtain the necessary knowledge to develop approaches for creating a vaccine against ASF.</span></p><p class="MsoNormal"><strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Materials and methods</span></strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">. ASFV isolate Krasnodar 07/17 and strain ASF/ARRIAH/CV-1 were used. Cloning of X69R, EP402R, and E248R genes was performed in the pJET1.2 / blunt vector and pCI-neo in E. coli JM-109 cells, according to the manufacturer's manual. Localization of recombinant proteins in CV-1 cell line carried out by direct immunofluorescence reaction (DIF) using polyclonal antibodies conjugated to FITC.</span></p><p class="MsoNormal"><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">The ASF virus reproduction level was assessed by hemadsorption reaction and qPCR kit (Central Research Institute of Epidemiology).</span></p><p class="MsoNormal"><strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Results</span></strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">. Recombinant plasmids pCI-neo / E248R, pCI-neo / EP402R and pCI-neo / X69R were constructed.</span></p><p class="MsoNormal"><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">The localization and the specificity of the obtained recombinant proteins CD2v, pE248R and pX69R was confirmed. It was established that these recombinant proteins induce the level of ASF virus reproduction on days 3-5 of the experiment by ~ 1.2-1.5 lgHADU<sub>50</sub>/cm<sup>3</sup> in comparison with the negative control.</span></p><p class="MsoNormal"><strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Discussion</span></strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">. The data obtained demonstrate the important role of CD2v, pX69R and pE248R proteins in the reproduction of the virus, since they significantly affect its level. The exact function of pX69R protein was not determined, however, in the experiments its positive effect on ASF virus reproduction was established, manifested in an increase in its reproduction level.</span></p><p class="MsoNormal"><strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">Conclusion</span></strong><span style="font-size: 14.0pt; line-height: 115%; mso-ansi-language: EN-US;" lang="EN-US">. This methodology allows us to study the nature of the effect of proteins with unknown function on ASF virus replication.</span></p>african swine fever virusrecombinant proteinslevel of viral accumulationvirus reproductionвирус африканской чумы свинейрекомбинантные белкиуровень накопления вирусарепродукция вируса[1. Garner G., Saville P., Fediavsky A., eds. African swine fever. Available at: http://lrd.spc.int/ext/Disease_Manual_Final/a120african_swine_fever.html][2. Rowlands R.J., Michaud V., Heath L., Hutchings G., Oura C., Vosloo W., et al. 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