复制型埃博拉病毒样颗粒的表达和鉴定

发布时间：2018-06-24 09:34

本文选题：埃博拉病毒 + 复制型病毒样颗粒　；参考：《病毒学报》2017年03期

【摘要】：为了研究埃博拉病毒的生物学特性及其与宿主的相互作用,构建无致病基因的复制型埃博拉病毒样颗粒(Transcription and replication competent virus-like particles,trVLPs)成为在BSL-2实验室中进行研究的理想途径。本研究目的是表达和鉴定复制型埃博拉病毒样颗粒(trVLPs)。本研究利用埃博拉病毒7个蛋白的基因组片段(VP40、VP24、GP、VP35、VP30、L、NP)和一个报告基因片段重组构建了表达质粒,瞬时转染293T细胞,通过荧光素酶检测系统测定报告基因的表达活性确定了复制型埃博拉病毒样颗粒(trVLPs)的成功表达。进一步通过超离转染细胞的上清纯化得到病毒样颗粒(trVLPs),在透射电子显微镜下观察到了典型的丝状结构。对病毒样颗粒进行荧光标记并感染Vero细胞,在荧光显微镜下观察到细胞中出现点状和丝状的荧光信号。复制型埃博拉病毒样颗粒的成功表达为进一步研究埃博拉病毒的生物学特性以及与宿主相互作用机制提供了基础。
[Abstract]:In order to study the biological characteristics of Ebola virus and its interaction with host, the construction of replication and replication competent virus-like particles-like VLPs with no pathogenic gene is an ideal way to study in BSL-2 laboratory. The aim of this study was to express and identify replicative Ebola virus-like particles (trVLPs). In this study, the expression plasmid was constructed using the genomic fragment of 7 proteins of Ebola virus (VP40, VP24GPG, VP35, VP30, LNP) and a reporter gene fragment, which was transiently transfected into 293T cells. The successful expression of replicative Ebola virus-like particles (trVLPs) was determined by luciferase detection system. Virus like particles (trVLPs) were further purified from the supernatant of the transfected cells. The typical filamentous structures were observed under transmission electron microscope (TEM). The virus like particles were labeled with fluorescence and infected with Vero cells. The fluorescence signals of dot and filamentous were observed under fluorescence microscope. The successful expression of replicative Ebola virus-like particles provides a basis for further studying the biological characteristics of Ebola virus and the mechanism of interaction with host.
【作者单位】：中国疾病预防控制中心病毒病预防控制所;中国科学院武汉病毒研究所;
【基金】：国家自然科学基金(项目号:81590763),题目:埃博拉病原学研究
【分类号】：R373

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