为构建高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)与致弱PRRSV的重组病毒,本研究在PRRSV致弱株APRRS的全长感染性克隆pAPRRS与HP-PRRSV感染性克隆pJX143的基础上,利用SOE-PCR技术将强弱毒的nsp2区域进行互换,构建了nsp2基因替换的强弱毒PRRSV嵌合感染性克隆.将构建的嵌合克隆转染MA104细胞,4d后观察到典型的细胞病变.通过RT-PCR和间接免疫荧光证明获得了强弱病毒株之间nsp2基因互换的嵌合病毒.这些嵌合病毒的构建和相应反向遗传操作平台的建立,为进一步研究HP-PRRSV的致病机制及相关疫苗的研发奠定了基础.%In order to construct recombinant virus between the highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) and the attenuated virus strain, based on an infectious cDNA clone of an attenuated PRRSV strain pAPRRS and the HP-PRRSV cDNA clone pJX143, we replaced the coding sequence of pAPRRS nsp2 with those of the HP-PRRSV by SOE-PCR. Upon transfection of recombinant virus into MA104 cells, typical PRRSV cytopathic effects were observed at 4 day post transfection. The rescued chimeric PRRSVs were identified by RT-PCR and indirect immunofluorescence assay. This study provided a valuable tool to develop the chimeric PRRSV as vaccine candidate offering cross-protection to HP-PRRSVs and to investigate the mechanism of pathogenesis of the increasing-virulence of the on-going prevalent PRRSV in China.
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