Identification of the Borna disease virus (BDV) proteins required for the formation of BDV-like particles.
Identification of the Borna disease virus (BDV) proteins required for the formation of BDV-like particles.
复制标题
鉴定形成 BDV 样颗粒所需的博尔纳病病毒 (BDV) 蛋白。
DOI：
10.1099/vir.0.80935-0
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
delaTorre,JuanCarlos
中科院分区：
文献类型：
--
作者：
Perez,Mar;delaTorre,JuanCarlos
博尔纳病病毒（Borna disease virus，BDV）是一种具有非节段负链RNA基因组的包膜病毒，其具有单负链病毒目的组织特征。然而，基于其独特的遗传学和生物学特性，BDV被认为是一个新的病毒家族Bornaviridae的原型成员。在此，描述了使用反向遗传方法来鉴定将BDV RNA类似物（MG）包装成感染性病毒样颗粒（VLP）所需的病毒蛋白。将在RNA聚合酶II启动子控制下编码单个BDV蛋白的质粒与允许由细胞RNA聚合酶I介导的BDV MG细胞内合成的质粒共转染。将来自转染细胞的澄清裂解物传代到新鲜细胞上，所述新鲜细胞先前用表达由BDV聚合酶介导的RNA合成所需的最小BDV反式作用因子L、N和P的质粒转染。通过存在于BDV MG中的氯霉素乙酰转移酶报告基因的表达来监测BDV MG特异性包装的重建和感染性VLP的传代。除L、N和P外，BDV M和G足以使氯霉素乙酰转移酶活性通过，该活性可被BDV针对G的中和抗体阻断，表明VLP感染性完全由BDV G介导。通过省略M或G废除BDV MG的传代。
Borna disease virus (BDV) is an enveloped virus with a non-segmented, negative-strand RNA genome that has an organization characteristic of Mononegavirales. However, based on its unique genetics and biological features BDV is considered to be the prototypic member of a new virus family,Bornaviridae. Here, the use of a reverse genetic approach to identify the viral proteins required for packaging of BDV RNA analogues (MG) into infectious virus-like particles (VLPs) was described. Plasmids encoding individual BDV proteins under the control of a RNA polymerase II promoter were co-transfected with a plasmid that allows for intracellular synthesis of a BDV MG mediated by the cellular RNA polymerase I. Clarified lysates from transfected cells were passaged onto fresh cells that were previously transfected with plasmids expressing the minimal BDVtrans-acting factors L, N and P required for RNA synthesis mediated by the BDV polymerase. Reconstitution of BDV MG-specific packaging and passage of infectious VLP was monitored by expression of the chloramphenicol acetyl transferase reporter gene present in the BDV MG. BDV M and G, in addition to L, N and P, were sufficient for the passage of chloramphenicol acetyl transferase activity, which could be blocked by BDV neutralizing antibodies to G, indicating that VLP infectivity was fully mediated by BDV G. Passage of BDV MG was abrogated by omission of either M or G.