表达猪圆环病毒2型ORF2基因的重组脑心肌炎病毒的构建与鉴定

发布时间：2018-04-30 19:14

本文选题：猪脑心肌炎病毒 + 猪圆环病毒型　；参考：《病毒学报》2017年02期

【摘要】：以脑心肌炎病毒(Encephalomyocarditis virus,EMCV)作为病毒载体,寻找基因组里理想的外源基因插入位点,为构建多价疫苗奠定基础。猪圆环病毒2型(Porcine circovirus type 2,PCV2)感染可引起断奶仔猪多系统衰竭综合征等多种疾病。脑心肌炎病毒(Encephalomyocarditis virus,EMCV)宿主谱广,能够天然感染多种哺乳动物。本研究采用EMCV NJ08毒株cDNA感染性克隆,将PCV2ORF2基因插入EMCV L蛋白基因第6位及第7位氨基酸之间,拯救获得重组EMCV rNJ08/Cap,拯救病毒含有分子遗传标记。Western Blot和IFA结果显示,该重组病毒成功表达Cap蛋白。重组病毒rNJ08/Cap在BHK-21细胞上生长曲线和病毒蚀斑大小与亲本病毒NJ08相似,证明EMCV可以作为表达外源基因的病毒载体,为PCV2和EMCV疫苗研究奠定了重要基础。
[Abstract]:Encephalomyocarditis virus (EMCV) was used as the virus vector to search for the ideal foreign gene insertion site in the genome and lay the foundation for the construction of multivalent vaccine. Porcine circovirus type 2 (PCV2) infection may cause multiple system failure syndrome in weaned piglets. Encephalitis virus Encephalomytis virusus (EMCV) has a wide host spectrum and can infect many mammals naturally. In this study, the cDNA infectious clone of EMCV NJ08 strain was used to insert PCV2ORF2 gene into the amino acids of EMCV L protein gene, and the recombinant EMCV rNJ08 / Cap. was obtained. The results of molecular genetic marker. Western Blot and IFA showed that the recombinant EMCV rNJ08 / Cap. The recombinant virus successfully expressed Cap protein. The growth curve of recombinant virus rNJ08/Cap on BHK-21 cells and the size of virus plaque were similar to those of parent virus NJ08, which indicated that EMCV could be used as a viral vector expressing exogenous gene, which laid an important foundation for the study of PCV2 and EMCV vaccines.
【作者单位】：南京农业大学农业部动物细菌学重点实验室;
【基金】：国家自然科学基金(项目号:31502082),题目:脑心肌炎病毒L蛋白影响I型干扰素信号通路的探索中央高校基本科研业务费专项资金(项目号:KJQN201616) 国家生猪产业技术体系专项(项目号:CARS-36)
【分类号】：S852.65

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