摘要
大菱鲆呼肠孤病毒(Scophthalmus maximus reovirus,SMRe V)属于呼肠孤病毒科水生呼肠孤病毒属水生呼肠孤病毒A型(AQRV-A),其病毒粒子具有双层衣壳,外衣壳由VP5和VP7蛋白构成。从SMRe V基因组中克隆出表达外衣壳蛋白VP5的全长基因vp5(2 057 bp),构建包含该基因全长的原核表达质粒p ET32a-vp5,转化E.coli表达菌BL21(DE3)。经诱导获得的融合蛋白以包涵体的形式表达,大小约为88 k D。将纯化的融合蛋白免疫小鼠,制备出抗SMRe V VP5的多抗血清。经Western blot杂交分析显示,该抗体制备成功,且能识别SMRe V的VP5蛋白,大小约为69 k D。进一步经间接免疫荧光分析显示,VP5呈颗粒状分布在宿主CIK细胞质中。
Scophthalmus maximus reovirus （SMReV） belongs to the Aquareovirus-A in the family Reoviridae. The virus particles are icosahedral in symmetry and have a double-layered capsid which is composed of VP5 protein and VP7 protein. The full-length （2 057 bp） ofvp5 was cloned from SMReV genome. The prokaryotic expression plasmid pET32a-vp5 was constructed and transformed into E.coli BL21 （DE3） in order to obtain recombinant protein. After being induced, the fusion protein was expressed as inclusion body with molecular weight of approximately 88 kD. The protein was purified and used to generate anti-SMReV VP5 sera in mice. Western blot analysis shows immune reaction of SMReV VP5 with sera, suggesting that the sera were successfully produced and recognized SMReV VP5 with molecular weight of about 69 kD, Indirect immunofluorescence assay （IFA） suggests that VP5 aggregated as granular structures in the cytoplasm of infected CIK cells.
出处
《南方水产科学》
CAS
CSCD
北大核心
2018年第1期43-49,共7页
South China Fisheries Science
基金
河南省自然科学基金面上项目(162300410041)
国家重点实验室开放课题(2016FB03)
河南科技大学博士科研启动基金(13480079)
