Loudon P T, Roy P
Laboratory of Molecular Biophysics, University of Oxford, United Kingdom.
Virology. 1992 Nov;191(1):231-6. doi: 10.1016/0042-6822(92)90184-q.
Bluetongue virus (BTV) core-like particles (CLPs) were synthesized by coexpression of VP3 and VP7 using a dual recombinant baculovirus. Purified CLPs were shown to bind single-stranded RNA in three different assay systems: gel retardation, nitrocellulose binding, and sucrose gradient sedimentation. CLPs showed equal affinity for BTV-specific and non-BTV RNA and also bound DNA. RNAase protection experiments demonstrated that bound RNA was accessible to immobilized ribonuclease, suggesting that the RNA was predominantly present on the outside of the CLPs. By using individually purified VP7 and VP3 in separate assays, the binding activity was shown to reside on VP3. These results indicate further functional homologies between BTV VP3 and the rotavirus inner-core VP2 protein.
使用双重组杆状病毒共表达VP3和VP7，合成了蓝舌病毒（BTV）核心样颗粒（CLP）。在三种不同的检测系统中，纯化的CLP显示出与单链RNA结合：凝胶阻滞、硝酸纤维素结合和蔗糖梯度沉降。CLP对BTV特异性RNA和非BTV RNA表现出同等亲和力，并且也能结合DNA。核糖核酸酶保护实验表明，固定化核糖核酸酶可接触到结合的RNA，这表明RNA主要存在于CLP的外部。通过在单独的检测中分别使用纯化的VP7和VP3，发现结合活性存在于VP3上。这些结果进一步表明BTV VP3与轮状病毒内核VP2蛋白之间存在功能同源性。
