减蛋下降综合征病毒重组Knob-S基因在杆状病毒系统中的表达及其免疫原性鉴定  

Expression of Knob-S gene of egg drop syndrome virus in baculovirus system and its immunogenicity

作　　者：郑金[1] 陈滔[1,2] 陆吉虎[2] 高峰[1,2] 候继波[2] 黎满香[1] 唐应华[2] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]江苏省农业科学院/国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《江苏农业学报》2015年第3期600-603,共4页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31201904);农业部公益性行业(农业)科研专项经费项目(201303046);科技部国家高技术研究发展计划"863"课题(2011AA10A213);江苏省农业科技自主创新基金项目[CX(14)5045]

摘　　要：为以杆状病毒系统在Sf9细胞中表达含减蛋下降综合征病毒(EDSV)重组Knob-S蛋白质来制备亚单位疫苗,采用PCR方法从鸭胚繁殖的EDSV尿囊液中扩增Knob-S基因,并克隆到p Fast Bac1中,经转化DH10Bac获得重组r Bac-Knob-S。将r Bac-Knob-S转染Sf9细胞获得重组杆状病毒,利用重组杆状病毒感染Sf9细胞表达重组蛋白质。重组Knob-S蛋白质经Western blotting和间接免疫荧光试验鉴定均为阳性。将重组Knob-S蛋白质制备成疫苗,按1羽0.3 ml(含50μg的重组病毒)免疫6周龄非免海兰褐雏鸡,在免疫后第2周可检测到血凝抑制抗体效价达8.5 lg2,在第3周达到高峰(9.5 lg2),表明重组Knob-S蛋白质具有较好的免疫原性。To develop egg drop syndrome virus subunit vaccine （ EDSV） based on its recombinant Knob-S proteins expressed in Sf9 cell, the Knob-S gene of EDSV was amplified from allantoic fluids of duck embryos by PCR, and cloned into pFastBac-1 before transformation into DH10Bac to construct rBac-Knob-S bacmid. The recombinant Knob-S proteins were expressed in Sf9 cell by infection of the recombinant baculovirus derived from the transfection of the bacmids in Sf9 cell. The recombinant Knob-S proteins were identified by Western blotting and indirect immunofluorescence assay. Groups of 6-week-old nonimmuned HY-Line brown chickens were inoculated with vaccines containing the recombinant Knob-S proteins. The serum antibody levels reached 8. 5 lg2 2 week post-vaccination, and rose to the peak of 9. 5 lg2 3 week post-vaccination determined by hemagglutinin inhibi-tion test, indicating that recombinant Knob-S proteins had good immunogenicity for chickens.

关 键 词：减蛋下降综合征病毒 重组Knob-S 真核表达 杆状病毒表达系统 

分 类 号：Q786[生物学—分子生物学]