I型猫冠状病毒的基因组组织与反向遗传分析
Genome organization and reverse genetic analysis of a type I feline coronavirus.
作者信息
Tekes Gergely, Hofmann-Lehmann Regina, Stallkamp Iris, Thiel Volker, Thiel Heinz-Jürgen
机构信息
Kantonal Hospital St. Gallen, Research Department, 9007 St. Gallen, Switzerland.
出版信息
J Virol. 2008 Feb;82(4):1851-9. doi: 10.1128/JVI.02339-07. Epub 2007 Dec 12.
Abstract
In this study we report the complete sequence and genome organization of the serotype I feline coronavirus (FCoV) strain Black. Furthermore, a reverse genetic system was established for this FCoV strain by cloning a full-length cDNA copy into vaccinia virus. This clone served as basis for the generation of recombinant FCoV (recFCoV) that was shown to bear the same features in vitro as the parental FCoV. Using this system, accessory 3abc genes in the FCoV genome were replaced by green fluorescent protein (recFCoV-GFP) and Renilla luciferase genes (recFCoV-RL). In addition, we showed that feline CD14(+) blood monocytes and dendritic cells can be easily detected after infection with recFCoV-GFP. Thus, our established reverse genetic system provides a suitable tool to study the molecular biology of serotype I FCoV.
摘要
在本研究中，我们报告了I型猫冠状病毒（FCoV）毒株Black的完整序列和基因组结构。此外，通过将全长cDNA拷贝克隆到痘苗病毒中，为该FCoV毒株建立了反向遗传系统。该克隆作为产生重组FCoV（recFCoV）的基础，重组FCoV在体外表现出与亲本FCoV相同的特征。利用该系统，FCoV基因组中的辅助3abc基因被绿色荧光蛋白（recFCoV-GFP）和海肾荧光素酶基因（recFCoV-RL）取代。此外，我们表明，用recFCoV-GFP感染后，可以很容易地检测到猫CD14(+)血液单核细胞和树突状细胞。因此，我们建立的反向遗传系统为研究I型FCoV的分子生物学提供了一个合适的工具。
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