Construction and biological characteristics analysis of a gene-deleted recombinant virus strain of LSDV-ORF151

文献类型： 外文期刊

第一作者： Li, Jiaqi

作者： Li, Jiaqi;Ke, Qunhua;Huang, Weitao;Ren, Shanhui;Li, Miaomiao;Yao, Kaishen;Ma, Xiaoqin;Sun, Yuefeng;Yin, Xiangping;Sun, Yuefeng

作者机构：

关键词： Lumpy skin disease virus; ORF151 gene; Gene deletion; Biological characterization; RNA-Seq

期刊名称：VIROLOGY （ 影响因子：2.4； 五年影响因子：2.5 ）

ISSN： 0042-6822

年卷期： 2025 年 610 卷

页码：

收录情况： SCI

摘要： Lumpy Skin Disease (LSD), caused by the Lumpy Skin Disease Virus (LSDV), is a highly virulent infectious disease that significantly impacts cattle health and economic productivity. The mechanisms underlying LSDV virulence and immune evasion remains poorly understood, and no commercial gene-deletion attenuated vaccine is currently available. This study aims to elucidate the functional role of LSDV genes and identify potential candidate strains for vaccine development. We focused on LSDV-ORF151, a gene potentially involved in immune response and apoptosis. Through amino acid sequence analysis and protein function prediction, we hypothesized that LSDV-ORF151 modulates host immune responses. RT-qPCR results showed that LSDV-ORF151 significantly reduces IFN-(3 transcription levels, suggesting its role in viral immune evasion. Using homologous recombination and limited dilution techniques, we constructed and purified a recombinant LSDV strain with an ORF151 deletion, using enhanced green fluorescent protein (EGFP) as a marker. PCR amplification and sequencing confirmed the stable inheritance of the rLSDV-zORF151-EGFP strain over at least 20 generations. Growth curve analysis revealed a slightly lower replication capacity compared to the wild-type LSDV strain (LSDV-WT), indicating that ORF151 deletion may attenuate viral replication. RT-qPCR showed that rLSDV-zORF151-EGFP induces higher IFN-(3 transcription levels than LSDV-WT after 24 h. Transcriptomic analysis indicated that the rLSDV-zORF151-EGFP strain induces heightened inflammatory and immune responses, and increased apoptosis in MDBK cells compared to LSDV-WT. This study provides a promising candidate for an LSDV gene-deletion attenuated vaccine and a theoretical foundation for further exploration of LSDV-ORF151's biological functions.

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