Abstract

African swine fever virus (ASFV) is producing a devastating pandemic that, since 2007, has spread to a contiguous geographical area from central Europe to Asia. In July 2021, ASFV was detected in the Dominican Republic, the first report of the disease in the Americas in more than 40 years. ASFV is a large, highly complex virus harboring a large dsDNA genome that encodes for more than 150 genes. The majority of these genes have not been functionally characterized. Bioinformatics analysis predicts that ASFV gene A859L encodes for an RNA helicase, although its function has not yet been experimentally assessed. Here, we evaluated the role of the A859L gene during virus replication in cell cultures and during infection in swine. For that purpose, a recombinant virus (ASFV-G-∆A859L) harboring a deletion of the A859L gene was developed using the highly virulent ASFV Georgia (ASFV-G) isolate as a template. Recombinant ASFV-G-∆A859L replicates in swine macrophage cultures as efficiently as the parental virus ASFV-G, demonstrating that the A859L gene is non-essential for ASFV replication. Experimental infection of domestic pigs demonstrated that ASFV-G-∆A859L replicates as efficiently and induces a clinical disease indistinguishable from that caused by the parental ASFV-G. These studies conclude that the predicted RNA helicase gene A859L is not involved in the processes of virus replication or disease production in swine.

Highlights

  • African swine fever (ASF) is currently affecting the swine production industry in a large geographical area stretching across Europe and into East and Southeast Asia

  • Recombinant mutant African swine fever virus (ASFV)-G-∆A859L was purified to homogeneity by successive rounds of limiting dilution purification based on mCherry activity detection. and was full-length sequenced using next-generation sequencing (NGS)

  • A859L Gene Is Conserved across Different ASFV Isolates

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Summary

Introduction

African swine fever (ASF) is currently affecting the swine production industry in a large geographical area stretching across Europe and into East and Southeast Asia. The development of ASFV experimental vaccines has focused on the deletion of specific virus genes involved in disease production. The resulting attenuated viruses have been shown effective in preventing disease during challenges with parental virulent strains [3,4,5,6,7] The development of these vaccines depends on the identification, characterization and genetic manipulation of virus genes involved in virus virulence. ASFV replication in swine macrophage cultures and during experimental infection in domestic pigs

Viruses and Cells
Detection of A859L Transcription
Construction of the ASFV A859L Deletion Mutant
Next-Generation Sequencing of ASFV
Animal Experiments
A859L Gene Is Conserved across Different ASFV Isolates
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