猪生殖与呼吸综合征病毒核衣壳蛋白在昆虫细胞中的表达及其用作诊断抗原的研究  

Expression of Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein in Insect Cells and Its Application for Diagnostic Antigen

作　　者：杜文金[1] 刘卫[2] 王志亮[1] 孙融冰[1] 宋翠平[1] 况慧星[2] 

机构地区：[1]农业部动物检疫所核心生物技术实验室,青岛266032 [2]新加坡国立大学分子农业生物学院,新加坡117604

出　　处：《中国预防兽医学报》2000年第S1期40-43,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：对猪生殖与呼吸综合征病毒（ＰＲＲＳＶ）核衣壳蛋白（Ｎ）作为ＥＬＩＳＡ抗原进行了研究，将编码ＰＲＲＳＶＮ蛋白的基因０ＲＦ７ｃＤＮＡ插入杆状病毒表达载体ｐＢｌｕｅＢａｃＨｉｓ２Ａ，通过同源重组获得了重组杆状病毒ＯＲＦ７－ＡｃＭＮＰＶ，感染昆虫细胞ＳＦ９表达了Ｎ蛋白，占细胞总蛋白的７．０７％，纯化后作为抗原建立了间接ＥＬＩＳＡ，与ＩＤＥＸＸ公司的ＥＬＩＳＡ诊断试剂盒有９６％的符合率，与ＩＦＡ有１００％的符合率。试验证实：ＰＲＲＳＶＮ蛋白在昆虫细胞中得到高效表达且是检测ＰＲＲＳＶ抗体的良好抗原。A novel indirect ELISA using recombinant porcine reproductive and respiratory syndrome virus nucleocapsid protein as antigen was developed. The cDNA of PRRSV ORF7, which encodes nucleocapsid protein, was inserted into baculovirus expression vector pBlue- His2A. Recombanant baculoviruses were gained. After infection of SF9 cells, the expressed N protein could reach up to 7 .07% of the total cell proteins and can be detected by Westerblat, IFA. The affinity -purified protein was used as ELISA antigen. ...

关 键 词：猪生殖和呼吸综合征病毒 重组核衣壳蛋白 酶联免疫吸附试验 

分 类 号：S85[农业科学—兽医学]