Expression of the outer capsid protein VP5 of two bluetongue viruses, and synthesis of chimeric double-shelled virus-like particles using combinations of recombinant baculoviruses.
Expression of the outer capsid protein VP5 of two bluetongue viruses, and synthesis of chimeric double-shelled virus-like particles using combinations of recombinant baculoviruses.
复制标题
两种蓝舌病毒外衣壳蛋白VP5的表达，以及使用重组杆状病毒组合合成嵌合双壳病毒样颗粒。
DOI：
10.1016/0042-6822(91)90620-q
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发表时间：
1991
期刊：
影响因子：
3.7
通讯作者：
Roy,P
中科院分区：
文献类型：
--
作者：
Loudon,PT;Hirasawa,T;Oldfield,S;Murphy,M;Roy,P
我们之前曾报道过在与重组杆状病毒共感染的草地贪夜蛾细胞中，由蓝舌病毒（BTV）的四种主要结构蛋白组成的病毒样颗粒（VLP）的组装（Frenchet al.（1990）.J. Virol.64，5695-5700）。在本文中，我们报告了进一步的研究，使用该系统组装异源VLP含有外部衣壳蛋白（VP 2和VP 5）的一系列不同的BTV血清型。用三种重组杆状病毒共感染frugiperdacells;表达VP 3和VP 7（分别为BTV-17和-10的）的双重重组体与表达BTV-1、-2、-10、-11、13或-17的VP 2的单一重组体以及表达BTV-2、BTV-10或BTV-13的VP 5的额外单一重组体组合。纯化所得的VLP，并通过电子显微镜、Western免疫印迹和血凝试验进行分析，以确定是否已组装双壳VLP。在这些实验的过程中，所有六种可用血清型的VP 2蛋白被成功地掺入VLP中。来自嵌合VLP的两种不同组合的颗粒（具有来自BTV-1或BTV-17的VP 2）用于在豚鼠中产生抗血清。这两种血清均显示出高的抗BTV活病毒中和抗体滴度，表明异源VLP可能具有用于抗BTV疫苗的潜力。
We have previously reported the assembly of virus-like particles (VLPs), consisting of the four major structural proteins of bluetongue virus (BTV), inSpodoptera frugiperdacells coinfected with recombinant baculoviruses (Frenchet al. (1990).J. Virol.64, 5695–5700). In this paper we report further studies using this system to assemble heterologous VLPs containing the outer capsid proteins (VP2 and VP5) of a range of different BTV serotypes.S. frugiperdacells were coinfected with three recombinant baculoviruses; a dual recombinant expressing VP3 and VP7 (of BTV-17 and −10, respectively) in combination with a single recombinant expressing VP2 of BTV-1, −2, −10, −11, 13, or −17 and an additional single recombinant expressing VP5 of BTV-2, BTV-10, or BTV-13. The resultant VLPs were purified and analyzed by electron microscopy, Western immunoblotting, and hemagglutination assays to determine whether double-shelled VLPs had been assembled. In the course of these experiments the VP2 proteins of all six available serotypes were successfully incorporated into VLPs. Particles from two different combinations of chimeric VLPs (having VP2 derived from BTV-1 or that of BTV-17) were used to raise antisera in guinea pigs. Both of these sera showed high neutralizing antibody titers against live BTV, indicating that heterologous VLPs may have potential for use in anti-BTV vaccines.