中国兽医科学2025，Vol.55Issue(11)：1437-1444,8.DOI:10.16656/j.issn.1673-4696.2025.0182
鸽Ⅰ型副黏病毒弱毒株构建及其生物学特性分析
Construction and biological analysis of an attenuated strain of pigeon paramyxovirus type 1
摘要
Abstract
The pigeon paramyxovirus type 1(PPMV-1)QY strain was attenuated through reverse genet-ics-mediated attenuation to generate a live-attenuated vaccine candidate targeting the predominant circulating genotype of PPMV-1.The full-length genome of the QY strain was partitioned into five discrete fragments(A-E)through enzymatic digestion at predetermined restriction enzyme recogni-tion sites.The genomic construct was engineered with a T7 promoter at the 5'-terminus and a hepatitis delta ribozyme(HDV Rz)exhibiting autocatalytic activity paired with a T7 terminator at the 3'-termi-nus.The F protein cleavage site was mutated to that of the La Sota strain.The amplified gene fragments were directionally cloned into the pOK-12X backbone vector to construct the recombinant plasmid desig-nated pOK-aQY.The NP,P,and L genes were subcloned into the pCI-neo expression vector to generate helper plasmids(pCI-NP,pCI-P,pCI-L),with functional validation performed through a minigenome re-porter assay in the reverse genetics system.These four plasmids were co-transfected into BSR-T7/5 cells to rescue the virus.The recombinant virus was validated through indirect immunofluorescence assay(IFA)and genetic marker analysis,followed by comprehensive characterization of genomic sta-bility and phenotypic properties in vitro.The recombinant plasmids pOK-aQY and helper plasmids(pCI-NP,pCI-P,pCI-L)were successfully generated,enabling rescue of the recombinant aQY strain through reverse genetics-mediated viral reconstitution.The aQY strain is stable,with an MDT of 108 h and an ICPI of 0.This study establishes reverse genetics as an effective attenuation platform for the QY strain,providing a molecular framework for elucidating PPMV-1 genomic determinants of pathogenici-ty and advancing genotype-specific vaccine development.关键词
鸽Ⅰ型副黏病毒/反向遗传学/病毒拯救/生物学特性/疫苗候选株Key words
pigeon paramyxovirus type 1/reverse genetics/virus rescue/biological characteris-tics/vaccine candidate分类
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陈佳薇,庞洪泽,雷白时,赵款,张云航,包银莉,蒋文明,张武超,袁万哲..鸽Ⅰ型副黏病毒弱毒株构建及其生物学特性分析[J].中国兽医科学,2025,55(11):1437-1444,8.基金项目
"十四五"国家重点研发计划项目(2023YFD1802602) （2023YFD1802602）
预防兽医学与生物技术福建省高等学校重点实验室开放基金项目(2021KF03) （2021KF03）
