中国兽医杂志2025，Vol.61Issue(9)：36-43,8.DOI:10.20157/j.cnki.zgsyzz.2025.09.005
伪狂犬病病毒US1缺失株的构建和生物学特性分析
Construction and Biological Characterization of US1 Gene-Deleted Pseudorabies Virus Strain
摘要
Abstract
Pseudorabies virus(PRV)is a member of the Alphaherpesvirinae family,and its unique short region1(US1)gene plays a critical role in the viral life cycle.However,the functional role of the US1 gene in PRV remains insufficiently studied.This study aimed to construct a US1 gene-deleted PRV strain(PRV-ΔUS1)and investigate its biological properties.First,a mouse-derived polyclonal antibody against PRV US1 protein was prepared through protein expression,purification,and immunization.The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9(CRISPR/Cas9)gene editing system was then used to generate the PRV-ΔUS1 strain,which was validated by polymerase chain reaction(PCR)and Western blot(WB)assays.Then,the biological characteristics of PRV-ΔUS1 were assessed using plaque assays,viral growth curves,and serial passage tests for genetic stability.The results showed successful generation of a high-titer mouse polyclonal antibody against PRV US1 protein.PCR and WB analysis confirmed the successful deletion of the US1 gene in PRV-ΔUS1.Compared with the wild-type strain,PRV-ΔUS1 exhibited significantly smaller plaque sizes(P<0.001)and markedly reduced replication capacity at 6,12,24,and 48 hours post-infection(P<0.001).Moreover,PRV-ΔUS1 maintained genetic stability over multiple passages.In conclusion,this study successfully constructed a US1 gene-deficient PRV strain and demonstrated that the US1 gene influences plaque formation and viral replication.These findings provide an important tool and theoretical basis for further functional studies of the US1 gene and the development of novel PRV vaccines.关键词
伪狂犬病病毒(PRV)/US1基因/CRISPR/Cas9/基因缺失病毒Key words
pseudorabies virus(PRV)/US1 gene/CRISPR/Cas9/gene-deleted virus分类
农业科技引用本文复制引用
屈玮钰,李鑫..伪狂犬病病毒US1缺失株的构建和生物学特性分析[J].中国兽医杂志,2025,61(9):36-43,8.基金项目
国家自然科学基金面上项目(32373020) （32373020）
