Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in China

Pseudorabies virus (PRV) is a highly contagious pathogen in swine that can cross species barriers and infect other mammals, including humans. Given the potential for interspecies transmission and its threat to public health, understanding the molecular biology of PRV strains is essential for develop...

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Main Authors: Mo Zhou, Haiyang Liang, Nannan Nie, Li Zhang, Rui Zhu, Shinuo Cao, Shanyuan Zhu
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Veterinary Sciences
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Online Access:https://www.mdpi.com/2306-7381/12/6/519
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author Mo Zhou
Haiyang Liang
Nannan Nie
Li Zhang
Rui Zhu
Shinuo Cao
Shanyuan Zhu
author_facet Mo Zhou
Haiyang Liang
Nannan Nie
Li Zhang
Rui Zhu
Shinuo Cao
Shanyuan Zhu
author_sort Mo Zhou
collection DOAJ
description Pseudorabies virus (PRV) is a highly contagious pathogen in swine that can cross species barriers and infect other mammals, including humans. Given the potential for interspecies transmission and its threat to public health, understanding the molecular biology of PRV strains is essential for developing effective control measures and preparing for future pandemics. In this study, a novel PRV strain, PRV-HL-2021, was isolated from an outbreak in Heilongjiang Province, China. The viral genome was used to establish a reverse genetics system based on a fosmid library of the PRV-HL-2021 genome. This system facilitated the creation of recombinant PRV, including one expressing EGFP and another with deletions in the <i>US9, gI,</i> and <i>gE</i> genes. PRV-HL-2021 was found to be highly lethal to mice <i>in vivo</i>. The recombinant PRV strains, such as rPRV-US9-EGFP and rPRV-del<i>gI/gE/US9</i>, exhibited growth characteristics similar to the parental PRV-HL-2021 strain. The isolation and characterization of PRV-HL-2021 contribute to a better understanding of the genetic diversity of PRV strains. The developed reverse genetics system provides valuable tools for investigating viral functions, creating genetically modified PRV strains, and advancing the development of safer vaccines. These findings will enhance strategies for controlling PRV outbreaks and mitigating its impact on both animal and public health.
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spelling doaj-art-2c5682e6c6f54d81a3489e682fc396c52025-08-20T03:32:31ZengMDPI AGVeterinary Sciences2306-73812025-05-0112651910.3390/vetsci12060519Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in ChinaMo Zhou0Haiyang Liang1Nannan Nie2Li Zhang3Rui Zhu4Shinuo Cao5Shanyuan Zhu6Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaNingbo Sansheng Biological Technology Co., Ltd., Ningbo 315000, ChinaMOA Key Laboratory of Animal Bacteriology, MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaPseudorabies virus (PRV) is a highly contagious pathogen in swine that can cross species barriers and infect other mammals, including humans. Given the potential for interspecies transmission and its threat to public health, understanding the molecular biology of PRV strains is essential for developing effective control measures and preparing for future pandemics. In this study, a novel PRV strain, PRV-HL-2021, was isolated from an outbreak in Heilongjiang Province, China. The viral genome was used to establish a reverse genetics system based on a fosmid library of the PRV-HL-2021 genome. This system facilitated the creation of recombinant PRV, including one expressing EGFP and another with deletions in the <i>US9, gI,</i> and <i>gE</i> genes. PRV-HL-2021 was found to be highly lethal to mice <i>in vivo</i>. The recombinant PRV strains, such as rPRV-US9-EGFP and rPRV-del<i>gI/gE/US9</i>, exhibited growth characteristics similar to the parental PRV-HL-2021 strain. The isolation and characterization of PRV-HL-2021 contribute to a better understanding of the genetic diversity of PRV strains. The developed reverse genetics system provides valuable tools for investigating viral functions, creating genetically modified PRV strains, and advancing the development of safer vaccines. These findings will enhance strategies for controlling PRV outbreaks and mitigating its impact on both animal and public health.https://www.mdpi.com/2306-7381/12/6/519pseudorabies virusfosmid librarygenetic manipulation platformrecombinant PRV
spellingShingle Mo Zhou
Haiyang Liang
Nannan Nie
Li Zhang
Rui Zhu
Shinuo Cao
Shanyuan Zhu
Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in China
Veterinary Sciences
pseudorabies virus
fosmid library
genetic manipulation platform
recombinant PRV
title Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in China
title_full Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in China
title_fullStr Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in China
title_full_unstemmed Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in China
title_short Identification, Pathogenicity, and Reverse Genetics System Construction of a Pseudorabies Virus Isolate from Pigs in China
title_sort identification pathogenicity and reverse genetics system construction of a pseudorabies virus isolate from pigs in china
topic pseudorabies virus
fosmid library
genetic manipulation platform
recombinant PRV
url https://www.mdpi.com/2306-7381/12/6/519
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