猪繁殖与呼吸综合征病毒XH株感染性克隆的构建  

Construction and identification of infectious clone for porcine reproductive and respiratory syndrome virus XH strain

作　　者：吴欣伟[1] 熊永忠[2] 秦宏阳[1] 曹宗喜[1] 于俊勇[1] 闫明菲[1] 郭泗虎[1] 张桂红[1] 

机构地区：[1]华南农业大学兽医学院农业部动物疫病防控重点开放实验室,广东广州510642 [2]中国农业科学院哈尔滨兽医研究所维科生物技术开发公司,黑龙江哈尔滨150069

出　　处：《中国预防兽医学报》2011年第2期97-100,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30871877);国家生猪现代农业产业技术体系(NYCYTX009)

摘　　要：为构建猪繁殖与呼吸综合征病毒(PRRSV)反向遗传操作系统,本实验采用RT-PCR方法将PRRSV-XH株的基因组序列分为6个片段进行扩增,连接至低拷贝载体pOKQ中,获得全长cDNA克隆。将该克隆体外转录,经脂质体转染Marc-145细胞,观察细胞病变。经RT-PCR、间接免疫荧光和western blot等试验验证,表明构建了具有感染性的PRRSV-XH株全长cDNA克隆。本实验为在分子水平上研究PRRSV的复制、致病机理和基因产物的功能等方面提供技术支持,并为构建新型病毒载体和开发安全有效的活病毒疫苗奠定基础。To establish a reverse genetic system of porcine reproductive and respiratory syndrome virus（PRRSV）,the complete genome of PRRSV XH strain was amplified by RT-PCR and cloned into low-copy vector pOKQ to construct PRRSV recombinant plasmid.RNA transcripts was produced by in vitro transcription and transfected into Marc-145 cells.Typical PRRSV CPE was developed following transfection.The rescued virus was identified by RT-PCR,IFA and western blot.The availability of PRRSV cDNA clone would provide a powerful means in the understanding of molecular mechanisms of PRRSV pathogenesis and the development of novel vaccines.

关 键 词：猪繁殖与呼吸综合征病毒 全长CDNA 拯救病毒 

分 类 号：S852.65[农业科学—基础兽医学]