传染性造血器官坏死病病毒糖蛋白杆状病毒表达系统的构建及其抗原性分析  

Construction of Baculovirus Expression System and Antigenicity Analysis of Infectious Hematopoietic Necrosis Virus Glycoprotein

作　　者：谢三磊[1] 温书香 罗琳[2] 马志宏[2] 李桂萍[1] 李铁梁[2] 姜娜[2] 邢薇[2] 孙惠玲[1] XIE San-lei WEN Shu-xiang LUO Lin MA Zhi-hong LI Gui-ping LI Tie-liang JIANG Na XING Wei SUN Hui-ling(Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China Beijing Fisheries Research Institute, Beijing 100068, China)

机构地区：[1]北京市农林科学院畜牧兽医研究所,北京畜禽疫病防控技术北京市重点实验室,北京100097 [2]北京市水产科学研究所,北京100068

出　　处：《中国畜牧兽医》2017年第3期854-860,共7页China Animal Husbandry & Veterinary Medicine

基　　金：北京市科委一般项目(Z141100002314002);北京市农林科学院科技创新能力建设专项(KJCX20140401)

摘　　要：为研究传染性造血器官坏死病病毒(infectious haematopoietic necrosis virus,IHNV)主要结构蛋白糖蛋白(G),本研究采用RT-PCR方法从IHNV中提取RNA并进行反转录,经PCR方法扩增获得1 380bp的G蛋白基因片段,将其克隆到pFB-LIC-Bse杆状病毒载体中,成功构建了重组质粒pFB-LIC-Bse-G,转化到大肠杆菌DH10Bac感受态细胞中,获得了重组杆粒rBacmid-G。将重组杆粒转染至Sf9昆虫细胞,获得了重组杆状病毒。间接免疫荧光(IFA)和Western blotting分析显示,重组G蛋白可与抗组氨酸单抗(Anti-His)、抗IHNV鼠血清发生特异性反应,表明本研究克隆的IHNV G蛋白在真核表达系统中得到正确表达,且该蛋白具有良好的抗原性。本试验结果为研究G蛋白的功能及开发IHNV新型疫苗奠定了基础。To study the major structural protein glycoprotein（G）of infectious hematopoietic necrosis virus（IHNV）,glycoprotein gene（1 380bp）was amplified by RT-PCR from IHNV.In order to construct a recombinant plasmid pFB-LIC-Bse-G,Ggene was cloned into the baculovirus vector pFB-LIC-Bse.Then,the constructed plasmid pFB-LIC-Bse-G was transformed into E.coli DH10 Bac.The recombinant bacmids rBacmid-G was got,and then it was transfected to insect Sf9 cells,and the recombinant baculovirus that contained Ggene was obtained.Western blotting analysis and indirect immunofluorescence assay（IFA）showed that the recombinant G protein could be recognized by histidine monoclonal antibody（Anti-His）and anti-IHNV antibody of mouse.The results indicated IHNV G protein had been expressed correctly in Sf9 cells.The study laid a foundation for further studying the G protein structure,function and immunological characteristics.

关 键 词：传染性造血器官坏死病病毒 糖蛋白 重组杆状病毒 

分 类 号：S941.41[农业科学—水产养殖]