Construction of Fluorescently Labelled Pseudorabies Viruses and Their Preliminary Application in Antiviral Drug Screening
In order to screen effective antiviral drugs against pseudorabies virus(PRV),we here constructed the recombinant strains PRV-mCherry expressing red fluorescent protein mCherry based on the human-originated isolate hSD-1/2019 and classical strain Ea.Specifically,mCherry reporter gene was inserted into the site before the terminator of PRV UL35 through homologous recombination technique.Based on the constructed recombinant fluorescent viruses,a high-throughput drug screening platform indicated by fluorescence intensity was established and ap-plied for testing 1 621 drugs in the natural product library for screening effective antiviral drugs.Properties of the drugs were preliminarily explored.The results showed that mCherry was in-serted in the correct position and was stably inherited.The fluorescence intensity of the recombi-nant strain PRV-mCherry was representative of the viral proliferation.The strains PRV-mCherry showed similar proliferation curves as the parental strains,supporting the usability of the PRV-mCherry strains in drug screening.Three drugs that could effectively inhibit PRV proliferation in vitro were screened out from the natural product library by using PRV-mCherry.Based on the determined 50％cytotoxic concentration and 50％inhibitory concentration,the selection indices of the three drugs were calculated to be 203.63-564.58 μmol·L-1,indicating that they had good drug-forming properties.The research provided new ideas in antiviral drugs for the treatment of PRV infection.
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