整合IBDV-VP_(2)基因的重组CVI988载体疫苗的构建  

Construction of recombinant CVI988 vector vaccine integrating IBDV-VP_(2)gene

作　　者：龚乐乐 黄鑫祥 康云哲 王乐乐[1] 邱祥琦 张圆圆[1] 高美洁 朱文慧 张雨林 庄国庆 孙爱军 GONG Lele;HUANG Xinxiang;KANG Yunzhe;WANG Lele;QIU Xiangqi;ZHANG Yuanyuan;GAO Meijie;ZHU Wenhui;ZHANG Yulin;ZHUANG Guoqing;SUN Aijun(International Joint Research Center of National Animal Immunology,College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学院国家动物免疫学国际联合研究中心,河南郑州450046

出　　处：《中国兽医学报》2024年第9期1865-1871,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(U21A20260);河南省高等学校重点科研基金资助项目(22A230011);河南农业大学高层次“拔尖人才”资助项目(30500690)。

摘　　要：传染性法氏囊病病毒(infectious bursal disease virus,IBDV)变异株的出现以及毒株的毒力增加对养鸡业构成严重威胁,构建表达IBDV-VP_(2)蛋白的CVI988重组候选疫苗株,可以同时防控IBDV和马立克病病毒(Marek’s disease virus,MDV)对养禽业造成的危害。本研究通过细菌人工染色体(BAC)技术,将IBDV-VP_(2)基因整合到马立克病(Marek’s disease,MD)疫苗株CVI988的UL55位点,得到重组病毒CVI988 BAC-VP_(2)。利用PCR,IFA对重组病毒进行鉴定,并对重组病毒的生物学特性进行研究。试验结果显示,重组病毒CVI988 BAC-VP_(2)成功拯救且能在鸡胚成纤维细胞(CEF)中稳定表达。重组病毒表现出与亲本病毒相似的体外复制能力,重组病毒的噬斑大小与亲本毒相比,差异不显著。本研究构建的重组病毒CVI988 BAC-VP_(2)将为研发同时防控IBDV和MDV感染的二联廉价疫苗奠定基础。The emergence of high virulent mutant strains of infectious bursal disease virus(IBDV)becomes a serious threat to the poultry industry.However,the live attenuated IBDV vaccine can potentially revert to a virulent strain.Therefore,it is a necessary to develop safe and effective IBDV-associated vaccines.The construction of a recombinant Marek's disease(MD)vaccine strain,CVI988,expressing the IBDV VP_(2)protein,can protect against disease induced by both IBDV and Marek's disease virus(MDV).Here,the IBDV-VP_(2)gene was integrated into the UL55 locus of CVI988 by bacterial artificial chromosome(BAC)technique,resulting in the recombinant virus CVI988 BAC-VP_(2).The recombinant virus was characterized by PCR,IFA and subsequently the biological properties of the recombinant virus were investigated.The results showed that the recombinant virus CVI988 BAC-VP_(2)was successfully rescued.The VP_(2)protein stably expressed in chicken embryo fibroblasts(CEF).The growth kinetics and plague size assays showed that there was comparable replication ability between recombinant virus and parental virus.This study provides the basis for the development of a low-cost vaccine against both IBDV and MDV infections.

关 键 词：细菌人工染色体 马立克病病毒 传染性法氏囊病病毒 活载体疫苗 

分 类 号：S858.31[农业科学—临床兽医学]