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  

Construction and Identification of a Recombinant EMCV Expressing ORF2 of Porcine Circovirus Type 2

作　　者：李亮[1] 范慧[1] 武迪[1] 王先炜[1] 姜平[1] 白娟[1] 

机构地区：[1]南京农业大学农业部动物细菌学重点实验室,江苏南京210095

出　　处：《病毒学报》2017年第2期224-230,共7页Chinese Journal of Virology

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)

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。We used the encephalomyocarditis virus（EMCV）as a viral vector to find the ideal insertion site of a＂foreign＂gene.This strategy lay the foundation for construction of a multivalent vaccine.Infection by the porcine circovirus type 2（PCV2）can cause various diseases,such as the post-weaning multisystemic wasting syndrome.We fused the PCV2ORF2 gene in the region of amino acids 6and 7in the leader protein gene of the EMCV NJ08 strain to generate a recombinant EMCV rNJ08/Cap-containing Cap gene.It was a special molecular marker.Results of western blotting and independent factor analysis showed that the recombinant virus expressed the Cap protein.The growth characteristics and plaque size of the recombinant virus rNJ08/Cap were similar to those of the parent virus NJ08.These findings suggested that EMCV could be used as a viral vector to express foreign genes.This is an important foundation for the development of vaccines against EMCV and PCV2.

关 键 词：猪脑心肌炎病毒 猪圆环病毒2型 病毒载体 

分 类 号：S852.65[农业科学—基础兽医学]