[轮状病毒的反向遗传学:仅从编码轮状病毒基因组的11个cdna中产生重组人轮状病毒]。

Uirusu Pub Date : 2019-01-01 DOI:10.2222/jsv.69.1
Satoshi Komoto, Saori Fukuda
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引用次数: 0

摘要

最近建立了一个完全基于质粒的动物轮状病毒反向遗传系统。我们在增加NSP2和NSP5基因cDNA质粒比例(3倍或5倍)的条件下,通过只转染11个T7质粒来生成重组轮状病毒(11质粒系统)。利用这种高效的系统,我们设计了第一个携带荧光(EGFP和mCherry)蛋白基因的传染性重组轮状病毒。除了这些重组动物病毒外,还利用11质粒系统进行了一些修饰,生成了第一个传染性重组人轮状病毒(KU (G1P[8])株)。重组人轮状病毒的可用性将为更好地了解这种医学上重要病毒的复制、致病性和其他生物学特性提供一个遗传学平台,并使下一代人轮状病毒疫苗的合理开发成为可能。
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[Reverse genetics of rotaviruses: Generation of recombinant human rotaviruses from just 11 cDNAs encoding the rotavirus genome].

An entirely plasmid-based reverse genetics system for animal rotavirus was established very recently. We improved the reverse genetics system to generate recombinant rotavirus by transfecting only 11 T7 plasmids for its 11 genes under the condition of increasing the ratio (3- or 5-fold) of the cDNA plasmids for NSP2 and NSP5 genes (11-plasmid system). Utilizing this highly efficient system, we engineered the first infectious recombinant rotaviruses harboring fluorescent (EGFP and mCherry) protein genes. In addition to these recombinant animal viruses, the first infectious recombinant human rotavirus (strain KU (G1P[8])) was also generated with the 11-plasmid system with some modifications. The availability of recombinant human rotaviruses will provide a genetic platform for a better understanding of the replication, pathogenicity, and other biological characteristics of this medically important virus and enable the rational development of next-generation human rotavirus vaccines.

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