中文摘要：

为构建表达猪繁殖与呼吸综合征病毒（PRRSV）GP5蛋白的重组犬2型腺病毒（CAV-2）,本研究将PRRSV GP5蛋白基因表达盒连接于E3区部分缺失的CAV-2全基因组重组质粒pPolyⅡ-CAV-2中,构建重组质粒pPoly-Ⅱ-CAV-2-GP5。并采用PmeⅠ和AscⅠ双酶切和脂质体介导法,将其转染MDCK细胞,获得重组病毒rCAV-2-GP5,经酶切鉴定显示该重组病毒含有目的基因。经RT-PCR检测表明,rCAV-2-GP5能够转录相应GP5蛋白基因mRNA;western blot检测证实GP5蛋白在MDCK细胞中得到表达。体外连续传30代检测表明,rCAV-2-GP5具有良好的遗传稳定性。本实验为PRRSV新型疫苗的研制奠定了基础。

英文摘要：

To construct the recombinant the canine adenovirus type-2（CAV-2） expressing the GP5 protein of porcine reproductive and respiratary syndrome virus（PRRSV）,a recombinant plasmid pPoly-Ⅱ-CAV-2-GP5 containing GP5 gene in the CAV-2 genome was generated.The CAV-2-GP5 genome sequence was obtained by restriction enzyme digestion and transfected into MDCK cells using Lipofectamine to generate the recombinant virus（rCAV-2-GP5）.rCAV-2-GP5 was screened by cytopathic effect and identified by restriction enzyme digestions,PCR,RT-PCR and western blot for GP5 protein expressing.The recombinant adenovirus showed genetic stability over 30 continuous passages.The rCAV-2-GP5 reported here could be used as a live virus vector to develop the vaccine against PRRSV.