家蚕幼虫中表达的登革热病毒血清型 2 的病毒样颗粒的形成。
Formation of Virus-Like Particles of the Dengue Virus Serotype 2 Expressed in Silkworm Larvae.
机构信息
Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, Shizuoka, 422-8529, Japan.
Laboratory of Biotechnology, Department of Agriculture, Graduate School of Integrated Science and Technology, Shizuoka University, Shizuoka, 422-8529, Japan.
出版信息
Mol Biotechnol. 2019 Nov;61(11):852-859. doi: 10.1007/s12033-019-00210-5.
To explore virus-like particles formation of dengue virus serotype type 2 (DENV-2) structural proteins of, C, prM, E were expressed in silkworm larvae using recombinant Bombyx mori nucleopolyhedroviruses (BmNPV). Each recombinant BmNPV bacmid coding the 2C-prM-E polypeptide and E protein fused with the signal peptide of bombyxin from B. mori was injected into silkworm larvae. The expressed proteins were collected from hemolymph and fat body, and purified using affinity chromatography. E protein was observed at 55 kDa. The DENV virus-like particles (DENV-LPs) with a diameter approximately 35 nm was observed using transmission electron microscopy (TEM) and immunogold-labelling TEM analysis. The binding of each partially purified proteins to heparin, one of receptors for DENV was confirmed. DENV-LPs were secreted in silkworm larval hemolymph even still low amount, but the E protein and heparin binding function were confirmed.
为了探索登革热病毒血清型 2（DENV-2）结构蛋白 2C、prM 和 E 形成类似病毒颗粒的情况，使用重组家蚕杆状病毒（BmNPV）在蚕幼虫中表达了这些蛋白。每个编码 2C-prM-E 多肽和与家蚕 bombyxin 信号肽融合的 E 蛋白的重组 BmNPV 杆状病毒亚基都被注射到蚕幼虫中。从血液和脂肪体中收集表达的蛋白，并通过亲和层析进行纯化。使用透射电子显微镜（TEM）和免疫金标记 TEM 分析观察到直径约为 35nm 的 DENV 样颗粒（DENV-LPs）。通过证实每种部分纯化蛋白与肝素（DENV 的一种受体）的结合，确认了 DENV-LPs 的存在。尽管量很少，但 DENV-LPs 仍在家蚕幼虫的血液中分泌，并且证实了 E 蛋白和肝素结合功能。
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