INFECTIOUS RABIES VIRUSES FROM CLONED CDNA
INFECTIOUS RABIES VIRUSES FROM CLONED CDNA
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DOI：
10.1002/j.1460-2075.1994.tb06739.x
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发表时间：
1994-09-15
期刊：
影响因子：
11.4
通讯作者：
CONZELMANN, KK
中科院分区：
文献类型：
--
作者：
SCHNELL, MJ;MEBATSION, T;CONZELMANN, KK
本文描述了完全由克隆的cDNA产生狂犬病毒（RV），这是一种横纹肌病毒科的非片段负链RNA病毒。在转染的质粒中，基因标记的全长RV抗基因组样T7 RNA聚合酶转录物和RV N、P和L蛋白在细胞内同时表达，导致转录活性核衣壳的形成和随后的传染性狂犬病病毒粒子的组装和出芽。除了真实的RV外，还从修饰的cDNA中恢复了两种具有预测转录模式特征的新型感染性RV。整个非翻译假基因区域的删除，在所有自然发生的rv中都是保守的，并不影响所产生的病毒在细胞培养中的繁殖。这表明非必要的遗传物质可能存在于非分段RNA病毒的基因组中。在另一种病毒的基因组中引入功能性的额外顺反子边界，导致转录额外的含有假基因序列的聚腺苷化mRNA。利用上述方法利用重组DNA技术操纵RV基因组的可能性——也可能适用于其他负链病毒——极大地促进了RV遗传学、病毒-宿主相互作用和狂犬病发病机制的研究，并为设计新一代活疫苗提供了工具。
The generation of infectious rabies virus (RV), a non-segmented negative-stranded RNA virus of the Rhabdoviridae family, entirely from cloned cDNA is described. Simultaneous intracellular expression of genetically marked full-length RV antigenome-like T7 RNA polymerase transcripts and RV N, P and L proteins from transfected plasmids resulted in formation of transcriptionally active nucleocapsids and subsequent assembly and budding of infectious rabies virions. In addition to authentic RV, two novel infectious RVs characterized by predicted transcription patterns were recovered from modified cDNA. Deletion of the entire non-translated pseudogene region, which is conserved in all naturally occurring RVs, did not impair propagation of the resulting virus in cell culture. This indicates that non-essential genetic material might be present in the genomes of non-segmented RNA viruses. The introduction of a functional extra cistron border into the genome of another virus resulted in the transcription of an additional polyadenylated mRNA containing pseudogene sequences. The possibility of manipulating the RV genome by recombinant DNA techniques using the described procedure-potentially applicable also for other negative-stranded viruses-greatly facilitates the investigation of RV genetics, virus-host interactions and rabies pathogenesis and provides a tool for the design of new generations of live vaccines.