Expression of rotavirus VP2 produces empty corelike particles.
Expression of rotavirus VP2 produces empty corelike particles.
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轮状病毒VP2的表达产生空的核状颗粒。
DOI：
10.1128/jvi.65.6.2946-2952.1991
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发表时间：
1991
影响因子：
5.4
通讯作者：
Cohen,J
中科院分区：
文献类型：
--
作者：
Labbe,M;Charpilienne,A;Crawford,SE;Estes,MK;Cohen,J
将牛轮状病毒RF株VP2全长基因克隆到杆状病毒转移载体pVL941的多角体启动子控制下。将野生型杆状病毒DNA与转移载体DNA共转染果蝇9细胞，得到含有牛轮状病毒基因2的重组杆状病毒。重组病毒感染果蝇细胞后，可产生与轮状病毒VP2相似的蛋白。该蛋白在重叠蛋白印迹实验中与双链RNA和DNA结合。表达的VP2以直径45 nm的芯样颗粒聚集在感染细胞的细胞质中。这些核状颗粒经蔗糖梯度离心法纯化后，发现不含核酸。异源轮状病毒(牛和猴)VP2和VP6的共表达导致了单壳颗粒的形成。这些结果明确地表明轮状病毒中存在一个独立于其他轮状病毒蛋白形成的最内层蛋白外壳。这些结果对轮状病毒的形态发生机制有一定的指导意义。
The complete VP2 gene of bovine rotavirus strain RF has been inserted into the baculovirus transfer vector pVL941 under the control of the polyhedrin promoter. Cotransfection of Spodoptera frugiperda 9 cells with wild-type baculovirus DNA and transfer vector DNA led to the formation of recombinant baculoviruses which contain bovine rotavirus gene 2. Infection of S. frugiperda cells with this recombinant virus resulted in the production of a protein similar in size and antigenic properties to the authentic rotavirus VP2. The protein binds double-stranded RNA and DNA in an overlay protein blot assay. Expressed VP2 assembles in the cytoplasm of infected cells in corelike particles 45 nm in diameter. These corelike particles were purified by sucrose gradient centrifugation and found to be devoid of nucleic acid. Coexpression of VP2 and VP6 from heterologous rotavirus strains (bovine and simian) resulted in the formation of single-shelled particles. These results definitively show the existence of an innermost protein shell in rotavirus which is formed independently of other rotavirus proteins. These results have implications for schemes of rotavirus morphogenesis.