EFFICIENT RECOVERY OF INFECTIOUS VESICULAR STOMATITIS-VIRUS ENTIRELY FROM CDNA CLONES
EFFICIENT RECOVERY OF INFECTIOUS VESICULAR STOMATITIS-VIRUS ENTIRELY FROM CDNA CLONES
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DOI：
10.1073/pnas.92.18.8388
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发表时间：
1995-08-29
影响因子：
11.1
通讯作者：
WERTZ, GTW
中科院分区：
文献类型：
--
作者：
WHELAN, SPJ;BALL, LA;WERTZ, GTW
从病毒基因组的全长cDNA克隆中回收了传染性水泡性口炎病毒（VSV）的原型非节段负链RNA病毒。使用从重组牛痘病毒表达的噬菌体T7 RNA聚合酶驱动在幼仓鼠肾细胞中从cDNA克隆合成VSV的基因组长度的正义转录物，所述幼仓鼠肾细胞同时表达VSV核衣壳蛋白、磷蛋白、通过噬斑测定，从10（6）个细胞的转染中获得多达10（5）个感染性病毒颗粒。该病毒在传代时扩增，被VSV特异性抗血清中和，并显示具有cDNA特征性的特异性核苷酸序列标记。这一成就使得VSV的生物学完全可用于病毒基因组的遗传操作，与正义RNA的成功相反，从负义T7转录物中回收感染性病毒的尝试都不成功，因为T7 RNA聚合酶在VSV基因间连接处或附近终止了转录。
Infectious vesicular stomatitis virus (VSV), the prototypic nonsegmented negative-strand RNA virus, was recovered from a full-length cDNA clone of the viral genome, Bacteriophage T7 RNA polymerase expressed from a recombinant vaccinia virus was used to drive the synthesis of a genome-length positive-sense transcript of VSV from a cDNA clone in baby hamster kidney cells that were simultaneously expressing the VSV nucleocapsid protein, phosphoprotein, and polymerase from separate plasmids, Up to 10(5) infectious virus particles were obtained from transfection of 10(6) cells, as determined by plaque assays, This virus was amplified on passage, neutralized by VSV-specific antiserum, and shown to possess specific nucleotide sequence markers characteristic of the cDNA, This achievement renders the biology of VSV fully accessible to genetic manipulation of the viral genome, In contrast to the success with positive-sense RNA, attempts to recover infectious virus from negative-sense T7 transcripts were uniformly unsuccessful, because T7 RNA polymerase terminated transcription at or near the VSV intergenic junctions.