表达PEDV S蛋白优势抗原区域的重组PRRSV活载体疫苗株的鉴定  

Characterization of a Recombinant PRRSV Live Vector Vaccine Strain Expressing the Dominant Antigenic Region of PEDV S Protein

作　　者：劳梦琴 刘瑞琳 陈鹏飞[1] 黄世静 于家荣 朱钧锐 孙祁 虞凌雪[1] 高飞[1] 姜一峰[1] 童武[1] 刘长龙[1] 李丽薇[1] 于海[1] 童光志[1] 周艳君[1] LAO Mengqin;LIU Ruilin;CHEN Pengfei;HUANG Shijing;YU Jiarong;ZHU Junrui;SUN Qi;YU Lingxue;GAO Fei;JIANG Yifeng;TONG Wu;LIU Changlong;LI Liwei;YU Hai;TONG Guangzhi;ZHOU Yanjun(Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]内蒙古农业大学兽医学院,呼和浩特010018

出　　处：《中国动物传染病学报》2025年第1期155-163,共9页Chinese Journal of Animal Infectious Diseases

基　　金：上海市科技兴农项目(2020-02-08-00-08-F01465);国家自然科学基金(32072861);上海市自然科学基金项目(20ZR1469600);国家生猪现代产业技术体系专项(CARS-35)。

摘　　要：猪繁殖与呼吸综合征(PRRS)和猪流行性腹泻(PED)是当前对养猪业影响较大的两种重要传染病,为了开发可同时针对这两种疫病的疫苗,本研究以PRRSV HuN4-F112活疫苗株为载体,将PEDV S蛋白中可诱导产生中和抗体的抗原区域进行优化和组合,命名为SNE,利用反向遗传操作技术将其引入至HuN4-F112株基因组中,构建了含有SNE的PRRSV全长cDNA克隆pHuN4-F112-SNE,通过病毒拯救获得了重组病毒rHuN4-F112-SNE株,经过对重组病毒引入基因和表达蛋白的鉴定,证实获得的重组病毒rHuN4-F112-SNE能够正确表达PEDV S蛋白优势抗原区域,其生物学特性与亲本毒株相似。将该重组病毒连续传至30代,引入的SNE基因均能获得稳定表达,且不影响亲本毒复制和自身蛋白的表达,表明重组病毒rHuN4-F112-SNE中引入的SNE基因可稳定遗传,研究结果为后续评价rHuN4-F112-SNE作为二价基因工程活载体疫苗的有效性奠定了基础。Porcine reproductive and respiratory syndrome(PRRS)and Porcine epidemic diarrhea(PED)are two important infectious diseases that currently have a great impact on the pig industry.In order to develop a vaccine that targeted both diseases at the same time,the live PRRSV HuN4-F112 vaccine strain was used in this study as the vector,the antigen region SNE in the PEDV S protein that induced neutralizing antibodies was inserted via reverse genetic manipulation technique.The PRRSV full-length cDNA clone pHuN4-F112-SNE was constructed and the recombinant virus rHuN4-F112-SNE strain was obtained through virus rescue.The resulting recombinant virus rHuN4-F112-SNE was confirmed and found to correctly express the dominant antigen region of PEDV S protein with similar biological characteristics to the parental strain.The SNE gene introduced into the recombinant virus was continuously expressed for 30 generations without affecting the replication and protein expression of the parental virus,indicating its stability.These results laid a foundation for future evaluation of the rHuN4-F112-SNE as a bivalent genetically engineered live vector vaccine.

关 键 词：PEDV S蛋白 PRRSV 重组病毒 

分 类 号：S859.797[农业科学—临床兽医学]