表达PRRSV NADC30-like毒株GP5-M的重组伪狂犬病病毒的构建及其生物学特性探究  

Construction and biological characteristics of a recombinant pseudorabies virus expressing GP5-M of PRRSV NADC30-like virus strain

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作  者:孙珊珊 其美拉姆 李强 曾南方 郑诚 张白玉 颜其贵[1] SUN Shanshan;CHEMI Lhamo;LI Qiang;ZENG Nanfang;ZHENG Cheng;ZHANG Baiyu;YAN Qigui(College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China;Shangchayu Town Agricultural and Animal Husbandry Comprehensive Center,ZayüCounty,Zayü860614,Xizang,China;Juxing Agriculture and Animal Husbandry Co.,Ltd.,Chengdu 611130,China;Sichuan Litian Animal Husbandry Co.,Ltd.,Dazhou 635099,Sichuan,China)

机构地区:[1]四川农业大学动物医学院,四川成都611130 [2]林芝市察隅县上察隅镇农牧综合中心,西藏察隅860614 [3]巨星农牧有限公司,四川成都611130 [4]四川省丽天牧业有限公司,四川达州635099

出  处:《浙江农业学报》2023年第11期2555-2567,共13页Acta Agriculturae Zhejiangensis

基  金:成都市重大科技应用项目(2022-YF09-00050-SN)。

摘  要:为获得一株能够融合表达猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)GP5-M蛋白的重组伪狂犬病病毒PRV-GP5-M毒株,以猪伪狂犬病病毒(pseudorabies virus,PRV)变异株TK基因缺失株PRV FJ01/TK-为病毒载体,以gI/gE基因为插入靶点,利用同源重组技术和CRISPR/Cas9技术敲除gI/gE基因,并在gI/gE位置上插入CMV-GP5-M表达盒,经噬斑纯化,成功构建能够正确表达GP5-M蛋白重组病毒PRV-GP5-M。进一步对该毒株的稳定性、生长动力学、培养特性、安全性等生物学特性进行探究。结果表明,该重组毒株具有良好的遗传稳定性、安全性,易于增殖培养。研究结果为靶向PRRSV GP5与M蛋白生成新的PRRSV疫苗提供了新的线索,同时可为预防近些年流行的PRRSV NADC30-like毒株和PRV变异株的疫苗研发提供重要参考。To obtain a recombinant pseudorabies virus PRV-GP5-M strain that can fuse and express the GP5-M protein of porcine reproductive and respiratory syndrome virus(PRRSV),using the porcine pseudorabies virus(PRV)variant TK gene deletion strain PRV FJ01/TK-as the viral vector and the gI/gE gene as the insertion target,the gI/gE gene was knocked out using homologous recombination technology and CRISPR/Cas9 technology,and a CMV-GP5-M expression box was inserted at the gI/gE position.After plaque purification,a recombinant virus PRV-GP5-M that could correctly express the GP5-M protein was successfully constructed.Furthermore,the biological characteristics of the strain,including stability,growth kinetics,culture characteristics and safety were explored.The results showed that the recombinant strain had good genetic stability,safety,and was easy to proliferate and cultivate.This study result provided new clues for targeting PRRSV GP5 and M protein to generate new PRRSV vaccines,and provided important references for vaccines against the prevalent PRRSV NADC30-like and PRV variant strains in recent years.

关 键 词:GP5-M蛋白 重组伪狂犬病病毒 CRISPR/Cas9 生物学特性 

分 类 号:S855.3[农业科学—临床兽医学]

 

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