伪狂犬病病毒荧光标签毒株构建及其在抗病毒药物筛选中的初步应用OA北大核心CSTPCD

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.