摘要
为了构建伪狂犬病病毒（PRV）TK/gI/gE三基因缺失重组病毒,试验以PRV-JL14作为母本株,将其基因组与转移载体进行同源重组后,以增强型绿色荧光蛋白基因（EGFP）作为标记基因进行筛选,先后敲除gI、gE和TK基因大部分序列,获得重组病毒PRV-ΔgIgE/TK,并将重组病毒在BHK-21细胞中传代培养并绘制其一步生长曲线,同时以不同剂量对小鼠进行攻毒试验。结果表明：TK、gI和gE基因缺失后的重组病毒基因组稳定,在BHK-21细胞中传代培养20代未见突变;与母本株PRV-JL14相比,PRV-Δg IgE/TK在BHK-21细胞中的增殖能力没有降低,且对小鼠的致病力显著降低。说明试验构建的重组病毒PRV-ΔgIgE/TK可以作为猪伪狂犬病的候选疫苗株。
The purpose of this study was to construct TK/gI/gE three gene-deleted recombinant pseudorabies virus（ PRV）. A PRV epidemic strain JL14 was used as a parental strain. After homologous recombination of its genome with the transfer vector,the screening was carried out by using the enhanced green fluorescent protein（ EGFP） gene as a marker gene. Then the recombinant virus PRV-ΔgIgE/TK was constructed by knocking out most sequence of the TK/gI/gE vene. The recombinant virus was passaged and cultured in BHK-21 cells and its one-step growth curve was drawn. At the same time,mice were challenged with different doses of recombiant virus. The results showed that the genome of the recombinant virus was stable after TK,gI and gE gene deletion,and no mutation was found in BHK-21 cells after being passaged for 20 generations. Compared with the parental strain,the proliferative ability of PRV-ΔgIgE/TK in BHK-21 cells did not decrease,but the pathogenicity of the recombinant virus to mice was significantly reduced. The results indicated that the recombinant virus PRV-ΔgIgE/TK could be used a candidate vaccine strain of swine pseudorabies.
作者
周鑫韬
刘晔
田宇飞
张艳艳
杨金金
阮晓凤
张守峰
扈荣良
ZHOU Xintao;LIU Ye;TIAN Yufei;ZHANG Yanyan;YANG Jinjin;RUAN Xiaofeng;ZHANG Shoufeng;HU Rongliang(College of Animal Seienee and Teehnology,Jilin Agricultural University,Changehun 130118,China;Military Veterinary Institute,Academy of Military Medical Sciences,Academy of Military Sciences,Changehun 130122,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2018年第17期13-17,25,230,共7页
Heilongjiang Animal Science And veterinary Medicine
基金
国家重点研发专项(2017YFD0500600)
