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.
出处
《中国兽医学报》
CAS
CSCD
北大核心
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)
