中国兽医科学2025，Vol.55Issue(6)：748-756,9.DOI:10.16656/j.issn.1673-4696.2025.0082
共表达IBDV VP2蛋白和AIV HA蛋白的重组NDV的构建
Construction of recombinant Newcastle disease virus co-expressing IBDV VP2 and AIV HA proteins
摘要
Abstract
To develop a trivalent live genotype Ⅶ Newcastle disease virus(NDV)vaccine against novel variant infectious bursal disease virus(nVarIBDV)and H9 subtype avian influenza virus(AIV),recombinant heat-resistant NDVs co-expressing nVarIBDV VP2 and AIV HA proteins were constructed.The nVarIBDV VP2 gene and AIV HA gene were linked by a 2A peptide,and cloned into the backbone of rNDV-Ⅶ-F to construct recombinant plasmids.Subsequently,the recombinant plasmid was co-transfected with the helper plasmid pTM-N/P/L into BHK-21 cells,enabling the rescue of recombinant viruses rNDV-Ⅶ-F-VP2-HA and rNDV-Ⅶ-F-HA-VP2,which co-express both the VP2 and HA proteins.RT-PCR confirmed that the target genes were successfully inserted between the P and M genes.IFA verified that both VP2 and HA proteins were stablely expressed in cell infected with rNDV-Ⅶ-F-HA-VP2,while only VP2 protein was detected in cell infected with rNDV-Ⅶ-F-VP2-HA.Biological characterization revealed that both recombinant viruses exhibited robust replication capacity while displaying attenuated phenotypes and comparable thermostability to the parental NDV strain.In conclusion,this study successfully constructed recombinant viruses rNDV-Ⅶ-F-VP2-HA and rNDV-Ⅶ-F-HA-VP2 that co-express the nVarIBDV VP2 protein and the H9N2 AIV HA protein,which provided a new strategy for the integrated prevention and control of nVarIBDV,H9N2 AIV,and geno-type Ⅶ NDV.关键词
新城疫病毒/传染性法氏囊病毒/VP2蛋白/禽流感病毒/HA蛋白Key words
Newcastle disease virus/infectious bursal disease virus/VP2 protein/avian influenza virus/HA protein分类
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王晓晓,何雷,司凯新,尚珂,陈松彪,余祖华,丁轲,陈建,李日顺,郁川..共表达IBDV VP2蛋白和AIV HA蛋白的重组NDV的构建[J].中国兽医科学,2025,55(6):748-756,9.基金项目
河南省科技攻关计划项目(222102110012,232102110078) （222102110012,232102110078）
