中国兽医科学2026，Vol.56Issue(9)：1148-1159,12.DOI:10.16656/j.issn.1673-4696.2026.0111
伪狂犬病病毒gI/gE/TK基因缺失疫苗株的构建及免疫效力评价
Construction and immunogenicity evaluation of a gI/gE/TK gene-deleted pseudorabies virus vaccine strain
摘要
Abstract
This study aimed to rapidly construct a thymidine kinase(TK)-deleted pseudorabies virus(PRV)using CRISPR/Cas9 technology.Using JS-2012-ΔgI/gE as the parental virus,two single-guide RNAs(sgRNAs)targeting the TK gene were designed and co-transfected with the viral genome into BHK-21 cells.The recombinant virus PRV JS-2012-ΔgI/gE/TK was obtained via plaque purification,PCR screen-ing and sequencing identification.Subsequently,in vitrobiological characterization,in vivo safety tests and efficacy tests were conducted.Biological characterization revealed that the recombinant virus had consistent in vitro growth characteristics and plaque morphology with the parental virus,lost TK gene activity,and exhibited good genetic stability.Animal test results showed that no mortal-ity occurred in mice within 8 days after inoculation with 1×106 TCID50 of the recombinant virus;piglets inoculated with 1×106 TCID50 of the strain remained safe within 14 days and could be effective-ly protected against challenge with 1×105 TCID50 of virulent PRV.This study provides a theoretical basis for the development of safer pseudorabies vaccines and disease eradication.关键词
伪狂犬病病毒/TK基因/CRISPR/Cas9/基因缺失Key words
pseudorabies virus/TK gene/CRISPR/Cas9/gene deletion分类
农业科技引用本文复制引用
孙依慧,姜一峰,于海,李丽薇,童光志,徐彦召,童武,李磊,刘元凯,王东丽,董一凡,周艳君,单同领,高飞,郑浩..伪狂犬病病毒gI/gE/TK基因缺失疫苗株的构建及免疫效力评价[J].中国兽医科学,2026,56(9):1148-1159,12.基金项目
国家重点研发计划项目(2022YFD1800800) （2022YFD1800800）
