表达猪繁殖与呼吸综合征病毒CH-1a株核衣壳蛋白重组鸡痘病毒的构建

从含有猪繁殖与呼吸综合征病毒（PRRSV）核衣壳蛋白（N）的质粒扩增出N基因，构建禽痘病毒转移载体。该载体含有禽痘病毒早晚期启动子LP2EP2控制之下的PRRSVN基因、P11启动下的报告基因lacZ以及用于同源重组的禽痘病毒基因组的片段。在转移载体转染亲本病毒S—FPV-017感染的鸡胚成纤维细胞（CEF）之后，采用蓝色表型筛选的方法，筛选到表达N基因的重组病毒，并对其进行了6轮蚀斑纯化。PCR方法鉴定证明重组病毒的基因组中含有完整PRRSVN基因，间接免疫荧光试验证明了PRRSVN蛋白在重组病毒感染的CEF细胞中获得表达，本研究为猪繁殖与呼吸综合征非复制型疫苗的研制打下了基础。

Construction of a recombinant fowlpox virus expressing nucleocapsid protein of porcine reproductive and respiratory syndrome virus

Nucleocapsid protein of porcine respiratory and reproductive virus was amplified, fowlpox virus transfer vector was constructed which contained fowpox virus promoter LP2EP2 controlled N gene, P11 controlled lacZ and homologous arm for recombination. S-FPV-017 infected chicken embryo fibroblast (CEF) cells were transfected with the transfer vector pSY681 N by lipofectin transfection procedure. The recombinant FPV (designated as rFPV-N) was screened by blue plague and purified by six passages, PCR results showed that N gene was inserted in the genome of rFPV-N. The expression of PRRSV N protein was tested by indirect immuno-fluorescence test, and the efficiency of the recombinant should be evaluated further.