Assembly of recombinant Israeli Acute Paralysis Virus capsids.

Assembly of recombinant Israeli Acute Paralysis Virus capsids.
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DOI:
10.1371/journal.pone.0105943
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发表时间:
2014
期刊:
影响因子:
3.7
通讯作者:
Jones IM
Jones IM
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Ren J;Cone A;Willmot R;Jones IM

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DicistroVirus以色列急性麻痹病毒(IAPV)与世界范围内蜜蜂数量的减少有关。IAPV和许多其他蜜蜂病毒在纯培养中的研究受到现有分离株和允许的细胞培养的限制。在这里,我们证明了将IAPV主要结构前体蛋白ORF2与其同源3C类处理酶偶联,导致在重组杆状病毒表达后,许多昆虫细胞系中单个结构蛋白的前体被加工。表达效率受IAPV 3C蛋白水平的影响,需要适度调节其活性才能获得最佳表达。成熟的IAPV结构蛋白组装成空衣壳,在蔗糖速度梯度上以颗粒的形式迁移,在电子显微镜下显示典型的DicistroVirus样形态。重组衣壳蛋白的单抗被配置成对IAPV存在的特异性诊断试验。微小核糖核酸病毒家族中的每一种蜜蜂病毒的重组衣壳都可能为正在进行的蜜蜂损失原因的调查提供病毒特异性试剂。
The dicistrovirus Israeli Acute Paralysis Virus (IAPV) has been implicated in the worldwide decline of honey bees. Studies of IAPV and many other bee viruses in pure culture are restricted by available isolates and permissive cell culture. Here we show that coupling the IAPV major structural precursor protein ORF2 to its cognate 3C-like processing enzyme results in processing of the precursor to the individual structural proteins in a number of insect cell lines following expression by a recombinant baculovirus. The efficiency of expression is influenced by the level of IAPV 3C protein and moderation of its activity is required for optimal expression. The mature IAPV structural proteins assembled into empty capsids that migrated as particles on sucrose velocity gradients and showed typical dicistrovirus like morphology when examined by electron microscopy. Monoclonal antibodies raised to recombinant capsids were configured into a diagnostic test specific for the presence of IAPV. Recombinant capsids for each of the many bee viruses within the picornavirus family may provide virus specific reagents for the on-going investigation of the causes of honeybee loss.
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发表时间: 2012
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影响因子: 3.7
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影响因子: 2.4
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