猪圆环病毒2型CAP蛋白在杆状病毒系统中的表达和鉴定  

Expression of porcine circovirus type 2 capsid protein using baculovirus system and identification of its serum reactivity

作　　者：冯向辉[1] 孙明[1] 刘伯华[1] 张丽[1] 乔明明[1] 申屠芬琴[1] 李智丽[1] 陈西钊[1,2] 

机构地区：[1]北京世纪元亨动物防疫技术有限公司,北京100085 [2]中国动物疫病预防控制中心,北京100125

出　　处：《中国兽医科学》2016年第2期223-227,共5页Chinese Veterinary Science

摘　　要：为更好地检测和防控猪圆环病毒2型(PCV2),利用bac PAK杆状病毒系统高效表达了PCV2 CAP蛋白。应用PCR扩增PCV2 ORF2基因,并将其克隆到载体pbac PAK9中,获得重组载体pbac PAK9-ORF2。用脂质体介导法将其与线性化杆状病毒bac PAK6共转染于Sf21细胞中,经蚀斑纯化获得纯的重组杆状病毒,并于优化表达条件后获得CAP蛋白。SDS-PAGE分析表明,CAP蛋白分子质量约为28 ku,表达量约为33 mg/L;间接ELISA试验表明,CAP蛋白与PCV2阳性血清产生特异性免疫反应,具有良好的反应原性。上述结果表明,本研究用杆状病毒系统高效表达了PCV2 CAP蛋白,为PCV2的分子诊断及亚单位疫苗的研制奠定了基础。To achieve high-level expression of porcine circovirus type 2（PCV2） CAP in vitro,the ORF2 gene of PCV2 was amplified and cloned into pbac PAK9 to construct the recombinant vector pbac PAK9-ORF2.The insect Sf21 cells were co-transfected with pbac PAK9-ORF2 and linearized bac PAK6. The recombinant baculovirus was obtained after plaque purification.Expression of CAP protein was analyzed by SDS-PAGE.The molecular weight of the expressed CAP is about 28 ku and the concentration of its production is about 33 mg/L. The successful usage of CAP as ELISA antigen proved its immunoreactivity. These results showed that the CAP protein produced in this study may be used for routine serodiagnosis of PCV2 infection and development of PCV2 subunit vaccine.

关 键 词：猪圆环病毒2型 重组杆状病毒 CAP蛋白 

分 类 号：S852.659.2[农业科学—基础兽医学]