摘要
根据1型鸭肝炎病毒E53株基因组序列设计并合成一对特异性引物,RT-PCR扩增VP1基因,将其定向克隆至pFastBacTMHTB载体,转化至DH10BacTM感受态细胞中,蓝白斑筛选获得重组穿梭质粒rBacmid-VP1,重组质粒在脂质体的介导下转染昆虫细胞Sf9,获得重组杆状病毒。结果表明VP1蛋白在昆虫细胞中获得表达,表达产物能够与兔抗1型鸭肝炎病毒VP1蛋白多抗血清和鸭肝炎病毒阳性血清发生特异性反应。在昆虫细胞中表达VP1蛋白为VP1功能的研究及鸭肝炎病毒抗体的检测提供了物质材料。
A pair of specific primers were designed based on genome subsequence of duck hepatitis virus type 1 E53, and VP1 gene was amplified by RT-PCR. The recombinant transfer vector pFastBacHTBTM-VP1 was constructed by cloning VP1 gene into the multiple clone site of baculovirus transfer vector pFastBacTMHTB and transformed into DH10BacTM competent cells for transposition. The correct recombinant bacmid was screened by blue/white spot selection, the isolated bacmid DNAs were transfected into Sf9 insect cells and packaged into baculovirus particles in the mediate of LipofectamineTM 2 000. The results showed that VP1 protein has been expressed in insect cells and the protein can react with polyclonal antibody and DHV-positive serum. The expression products of VP1 gene of duck hepatitis virus type 1 E53 will be an important substance for study on VP1 protein and the detection of antibodies against DHV.
出处
《中国家禽》
北大核心
2012年第8期24-27,共4页
China Poultry
基金
高等学校博士学科点专项科研基金资助课题(2010023225110004)
黑龙江省科技攻关项目(GB01B503-02
GB04B504)
东北农业大学博士启动基金
