河南农业科学2018，Vol.47Issue(1)：114-117,4.DOI:10.15933/j.cnki.1004-3268.2018.01.020
Ⅰ型鸭甲型肝炎病毒VP1基因在昆虫细胞中的表达及鉴定
Expression and Identification of VP1 Gene of Duck Hepatitis A Virus Type Ⅰ in Insect Cells
摘要
Abstract
In order to express the main structural protein VP1 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 VP1 gene by RT-PCR,the amplified fragment was cloned into baculovirus expression vector pFastBac1.The recombinant vector pFB-VP1 was transformed into E.coli DH10Bac competent cells,and the positive recombinant bacmid rBacmid-VP1 was screened according to the resistant and the blue-white plague screening,confirmed by PCR.The recombinant bacmid rBacmid-VP1 was transfected into the Sf 9 insect cells by liposome to obtain the recombinant baculovirus,and was named as rBac-VP1.The result of Western blot showed that the molecular weight of the recombinant proteins was about 27 ku,and could be recognized by the antiserum of DHAV-Ⅰ VP1.Indirect immunofluorescence analysis proved that the recombinant proteins could be recognized by the positive anti-virus serum.These results suggested that the main structural protein VP1 of DHAV-Ⅰ with immunogenicity was expressed successfully in insect cells.关键词
Ⅰ型鸭甲型肝炎病毒/VP1基因/昆虫细胞/表达/鉴定Key words
Duck hepatitis A virus type Ⅰ/VP1 gene/Insect cells/Expression/Identification分类
农业科技引用本文复制引用
顾玲玲,吴萌,朱善元,王安平..Ⅰ型鸭甲型肝炎病毒VP1基因在昆虫细胞中的表达及鉴定[J].河南农业科学,2018,47(1):114-117,4.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）
