摘要
采用RT-PCR方法从传染性造血器官坏死病病毒(IHNV)中扩增RNA获得1 176 bp的核衣壳蛋白(N)基因,将其插入于pFB-LIC-Bse杆状病毒载体中,构建了重组质粒pFB-LIC-Bse-N,转化至感受态细胞DH10Bac中,获得重组杆粒rBacmid-N。再将其转染至Sf9昆虫细胞,获得重组杆状病毒。IFA和Western Blot分析表明,重组N蛋白可以被辣根过氧化物酶(HRP)标记的组氨酸单抗(anti-His)识别,表明,IHNV N基因在昆虫细胞Sf9中获得了正确表达。为深入研究N蛋白的功能和免疫学特性奠定了基础。
To study the major structural nucleoprotein（ N） of infectious hematopoietic necrosis virus（ IHNV）by baculovirus systems,in this study nucleoprotein（ N） gene（ 1176 bp） was amplified by RT-PCR from infectious hematopoietic necrosis virus（ IHNV） and inserted into the baculovirus vector pFB-LIC-Bse to construct a recombinant plasmid pFB-LIC-Bse-N.The constructed recombinant transposition plasmid pFB-LIC-Bse-N was transformed to competent E.coli DH10Bac to get recombinant bacmids rBacmid-N.The obtained recombinant bacmid rBacmid-N was transfected to insect Sf9 cells,and the recombinant baculovirus that contained N gene was obtained.IFA and Western Blot analysis showed that the recombinant N protein can be recognized by horseradish peroxidase（ HRP）labeled histidine monoclonal antibody（ anti-His） which indicated IHNV N gene has been successfully expressed in Sf9 cells infected with recombinant baculovirus.The study laid a foundation of further study on N protein structure,function and immunological characteristics.
出处
《华北农学报》
CSCD
北大核心
2014年第3期36-40,共5页
Acta Agriculturae Boreali-Sinica
基金
北京市农林科学院科技创新基金(CXJJ201316)
