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  

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

作　　者：智海东[1] 王云峰[1] 张晶[1] 王玫[1] 刘娣 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]黑龙江省农业科学院博士后工作站,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2007年第1期1-4,26,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省博士后基金;国家高技术发展计划(863计划)(2001AA213041)的资助

摘　　要：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。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, P. 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.

关 键 词：猪繁殖与呼吸综合征病毒 核衣壳蛋白 重组鸡痘病毒 

分 类 号：S852.659.1[农业科学—基础兽医学]