Reverse genetics for peste-des-petits-ruminants virus (PPRV): Promoter and protein specificities
Reverse genetics for peste-des-petits-ruminants virus (PPRV): Promoter and protein specificities
复制标题
DOI：
10.1016/j.virusres.2007.01.015
复制
发表时间：
2007-06-01
期刊：
影响因子：
5
通讯作者：
Banyard, Ashley C.
中科院分区：
文献类型：
--
作者：
Bailey, Dalan;Chard, Louisa S.;Banyard, Ashley C.
小反刍兽疫病毒 (PPRV)（副粘病毒科，麻疹病毒属）会引起绵羊和山羊的急性发热性疾病，导致受感染牛群的显着发病率和死亡率。副粘病毒均具有阴性意义。非分段 RNA 基因组及其宿主范围和致病决定因素已使用反向遗传学进行了广泛研究。该技术还可以在疫苗设计方面采取更合理的方法。为了启动 PPRV 的此类工作，我们构建了 PPRV 微型基因组并研究了其在转染细胞中的表达。对于其他麻疹病毒，小基因组拯救的最低要求是基因组 (GP) 和反基因组 (AGP) 启动子的反式作用元件以及三种反式作用辅助蛋白 N（核衣壳）、P（磷蛋白）和 L（大聚合酶）。将同源 PPRV 辅助蛋白与密切相关的牛瘟病毒 (RPV) 的异源类似物进行比较，发现异源小基因组拯救的过程效率要低得多。通过设计两个 GP/AGP 嵌合小基因组，我们还鉴定了两种病毒在启动子和转录酶/复制酶复合物之间的特定相互作用方面的差异。 PPRV 小基因组在体外也被证明不严格遵守“六规则”。 (C) 2007 Elsevier B.V. 保留所有权利。
Peste-des-petits-ruminants virus (PPRV) (family Paramyxoviridae, genus Morbillivirus) causes an acute febrile illness in sheep and goats resulting in significant morbidity and mortality in infected herds. The paramyxoviruses all have negative sense. non-segmented RNA genomes and their host range and pathogenic determinants have been extensively studied using reverse genetics. This technology also enables a more rational approach to be taken with respect to vaccine design. In order to initiate this type of work for PPRV we constructed a PPRV minigenome and studied its expression in transfected cells. As for other morbilliviruses, the minimum requirements for minigenome rescue were shown to be the vis-acting elements of the genome (GP) and antigenome (AGP) promoters as well as the three trans-acting helper proteins N (nucleocapsid), P (phosphoprotein) and L (large polymerase). Homologous PPRV helper proteins were compared to their heterologous analogues from the closely related rinderpest virus (RPV) and heterologous minigenome rescue was found to be a much less efficient process. By engineering two GP/AGP chimeric minigenomes we also identified differences between the two viruses in the specific interactions between the promoters and the transcriptase/replicase complexes. The PPRV minigenome was also shown not to strictly comply with the "rule of six" in vitro. (C) 2007 Elsevier B.V. All rights reserved.