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[ASF virus replication features in the presence of recombinant proteins CD2v, pX69R and pE248R.]

A Mazloum¹, I U Zhukov¹, E B Aronova¹

¹FGBI «ARRIAH» Federal State Budgetary Institution «Federal Center for Animal Health» Vladimir region, Vladimir city, Yuryevets microdistrict, 600901, Russian Federation.

Voprosy Virusologii

|March 13, 2020

Abstract

Introduction:

African swine fever (ASF), sever hemorrhagic disease of swine caused by a large DNA virus of the Asfaviridae family. 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.

Purpose:

To study the effect of ASFV recombinant proteins CD2v, pE248R and pX69R on the speed and level of reproduction of ASF virus in vitro. Thus, obtain the necessary knowledge to develop approaches for creating a vaccine against ASF.

Materials And Methods:

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. The ASF virus reproduction level was assessed by hemadsorption reaction and qPCR kit (Central Research Institute of Epidemiology).

Results:

Recombinant plasmids pCI-neo / E248R, pCI-neo / EP402R and pCI-neo / X69R were constructed. 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 lgHADU₅₀/cm³ in comparison with the negative control.

Discussion:

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.

Conclusion:

This methodology allows us to study the nature of the effect of proteins with unknown function on ASF virus replication.

Keywords:

african swine fever virus level of viral accumulation recombinant proteins virus reproduction

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