Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening
Porcine reproductive and respiratory syndrome virus (PRRSV) is a significant pathogen posed a serious threat to the global swine industry. In this study, a BAC-based reverse genetics platform was established using a highly pathogenic PRRSV (HP-PRRSV) strain. Three recombinant reporter viruses were c...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2025-08-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1653170/full |
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| author | Rongxiao Liu Shi Fu Yukun Chai Nian Liu Benjin Liu Lingzhi Luo Wei Yang Xiumei Dong Jin Cui |
| author_facet | Rongxiao Liu Shi Fu Yukun Chai Nian Liu Benjin Liu Lingzhi Luo Wei Yang Xiumei Dong Jin Cui |
| author_sort | Rongxiao Liu |
| collection | DOAJ |
| description | Porcine reproductive and respiratory syndrome virus (PRRSV) is a significant pathogen posed a serious threat to the global swine industry. In this study, a BAC-based reverse genetics platform was established using a highly pathogenic PRRSV (HP-PRRSV) strain. Three recombinant reporter viruses were constructed by inserting the enhanced green fluorescent protein (EGFP) gene into three different intergenic regions of the complete PRRSV-L251 genome. Immunofluorescence assays combined with viral growth kinetics and reporter gene stability assessments indicated that rL251-ORF4-5a-EGFP maintained relatively stable expression during serial passage, and viral titers at 72 hours post-infection (hpi) were comparable to the parental virus. Subsequently, we identified four candidate compounds with potential anti-PRRSV activity using rPRRSV-L251-ORF4-5a-EGFP, indicating that this platform can be used as a visual assessment tool for antiviral drug screening. This study demonstrated that the ORF4-5a interval region is a feasible and promising site for exogenous gene insertion, and provided a robust technical platform for PRRSV vaccine development and pathogenesis studies. |
| format | Article |
| id | doaj-art-0bc8f45acb9b4d29a8e5edcd1a54b6ff |
| institution | Kabale University |
| issn | 2235-2988 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj-art-0bc8f45acb9b4d29a8e5edcd1a54b6ff2025-08-26T05:27:50ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-08-011510.3389/fcimb.2025.16531701653170Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screeningRongxiao LiuShi FuYukun ChaiNian LiuBenjin LiuLingzhi LuoWei YangXiumei DongJin CuiPorcine reproductive and respiratory syndrome virus (PRRSV) is a significant pathogen posed a serious threat to the global swine industry. In this study, a BAC-based reverse genetics platform was established using a highly pathogenic PRRSV (HP-PRRSV) strain. Three recombinant reporter viruses were constructed by inserting the enhanced green fluorescent protein (EGFP) gene into three different intergenic regions of the complete PRRSV-L251 genome. Immunofluorescence assays combined with viral growth kinetics and reporter gene stability assessments indicated that rL251-ORF4-5a-EGFP maintained relatively stable expression during serial passage, and viral titers at 72 hours post-infection (hpi) were comparable to the parental virus. Subsequently, we identified four candidate compounds with potential anti-PRRSV activity using rPRRSV-L251-ORF4-5a-EGFP, indicating that this platform can be used as a visual assessment tool for antiviral drug screening. This study demonstrated that the ORF4-5a interval region is a feasible and promising site for exogenous gene insertion, and provided a robust technical platform for PRRSV vaccine development and pathogenesis studies.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1653170/fullPRRSVBACEGFPantiviral drug screeningreverse genetics system |
| spellingShingle | Rongxiao Liu Shi Fu Yukun Chai Nian Liu Benjin Liu Lingzhi Luo Wei Yang Xiumei Dong Jin Cui Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening Frontiers in Cellular and Infection Microbiology PRRSV BAC EGFP antiviral drug screening reverse genetics system |
| title | Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening |
| title_full | Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening |
| title_fullStr | Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening |
| title_full_unstemmed | Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening |
| title_short | Construction of an EGFP-embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening |
| title_sort | construction of an egfp embedded porcine reproductive and respiratory syndrome virus infectious clone and antiviral drug screening |
| topic | PRRSV BAC EGFP antiviral drug screening reverse genetics system |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1653170/full |
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