四联重组痘苗病毒的构建及筛选  

Construction and selection of recombinant vaccinia virus with four genes combined

作　　者：王宏宇[1,2] 徐一鸣[3] 盛媛[1,2] 李一权 阎富龙[1,2] 赵权[1] 李霄[2] 金宁一[1,2] 

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]军事医学科学院军事兽医研究所,吉林长春130122 [3]吉林省兽药饲料监察所,吉林长春130062

出　　处：《中国兽医学报》2014年第9期1411-1415,1417,共6页Chinese Journal of Veterinary Science

基　　金：吉林省重点科技攻关资助项目(20130206041NY)

摘　　要：构建含黄热病毒E基因、汉坦病毒S基因、克里米亚-刚果出血热病毒M基因及炭疽杆菌POAX2基因的重组天坛株痘苗病毒。设计合成含外源筛选标记EGFP基因和4种拟插入外源基因的重组质粒,利用同源重组和荧光标记筛选技术并结合Cre/Loxp敲除系统,最终构建四联重组痘苗病毒。应用PCR对获得的重组痘苗病毒进行鉴定和遗传稳定性的检测,并在电子显微镜下观察其形态特征。构建含4种外源基因的重组痘苗病毒rVTT-C-4+,该重组痘苗病毒在转录水平上具有良好的遗传稳定性,且于电镜下观察具备完整形态。成功构建四联重组天坛株痘苗病毒,为后续疫苗研究奠定了基础。To construct a recombinant vaccinia virus contained Yellow fever virus envelope protein E gene, Hantaan virus S gene,Crimean-Congo hemorrhagic fever virus M gene and the POAX2 gene of Bacillus anthracis strain. We designed and synthesized a recombinant plasmid including four exogenous genes and EGFP gene, using homologous recombination combined with Cre/Loxp system to get recombinant vaccinia virus. Then,we used PCR to identify and detect genetic stabili- ty of the recombinant virus. At the same time, we observed the morphological characteristics of the recombinant virus by electron microscope observation. We got a recombinant vaccinia virus contained four exogenous genes. The recombinant vaccinia virus has a good genetic stability on transcription,and complete morphology. We successfully constructed the recombinant vaccinia vi- rus with four exogenous genes inserted and laid the foundation for subsequent research.

关 键 词：天坛株痘苗病毒 多联疫苗 EGFP Cre/Loxp敲除系统 

分 类 号：S852.65[农业科学—基础兽医学]