河南农业大学学报2017，Vol.51Issue(3)：365-369,5.
Ⅰ型鸭甲型肝炎病毒VP0基因在杆状病毒表达系统中的表达
Expression of VP0 genes of duck hepatitis A virus type-Ⅰ in insect cells
摘要
Abstract
In order to express the structural protein VP0 of duck hepatitis A virus type-Ⅰ (DHAV-Ⅰ) in insect cells,one pair of specific primers were designed according to the published genome sequences of DHAV-Ⅰ to amplify VP0 genes by PCR,the amplified fragment was cloned into Baculovirus expression vector pFastBac1.The recombinant vector pFB-VP0 was transformed into DH10Bac E.coli,and the positive recombinant bacmid rBacmid-VP0 was screened according to the resistant and the blue-white plague screening.The recombinant bacmid rBacmid-VP0 was transfected into the Sf9 insect cells by liposome.Once the cytopathic effect was found,the rBac-VP0 was aquired.The result of Western blot showed that the molecular weight of the recombinant proteins was about 29 kD.Indirect immunofluorescence analysis showed that the recombinant proteins could be recognized by the positive anti-virus serum.These results suggest that the structural proteins VP0 of DHAV-Ⅰ have been expressed successfully in insect cells,which can lay the foundation of function study on VP0 protein.关键词
Ⅰ型鸭甲型肝炎病毒/VP0基因/昆虫细胞/表达Key words
duck hepatitis A virus type-Ⅰ/VP0 gene/insect cells/expression分类
农业科技引用本文复制引用
王安平,顾玲玲,王永娟,吴双,左伟勇,洪伟鸣,朱善元..Ⅰ型鸭甲型肝炎病毒VP0基因在杆状病毒表达系统中的表达[J].河南农业大学学报,2017,51(3):365-369,5.基金项目
国家自然科学基金项目(31302096) （31302096）
江苏省农业支撑项目(BE2013415) （BE2013415）
江苏省六大人才高峰项目(NY-009) （NY-009）
