利用痘苗病毒载体进行冠状病毒的反向遗传学研究。
Reverse genetics of coronaviruses using vaccinia virus vectors.
作者信息
Thiel V, Siddell S G
机构信息
Research Department, Cantonal Hospital St Gallen, St Gallen, Switzerland.
出版信息
Curr Top Microbiol Immunol. 2005;287:199-227. doi: 10.1007/3-540-26765-4_7.
Abstract
In this article, we describe the reverse genetic system that is based on the use of vaccinia virus cloning vectors. This system represents a generic approach to coronavirus reverse genetics and was first described for the generation of recombinant human coronavirus 229E representing a group I coronavirus. Subsequently, the same approach has been used to generate recombinant avian infectious bronchitis coronavirus and, recently, recombinant mouse hepatitis virus, representing group III and group II coronaviruses, respectively. We describe how vaccinia virus-mediated homologous recombination can be used to introduce specific mutations into the coronavirus genomic cDNA during its propagation in vaccinia virus and how recombinant coronaviruses can be isolated. Finally, we describe how the coronavirus reverse genetic system has now been extended to the generation of coronavirus replicon RNAs.
摘要
在本文中，我们描述了基于痘苗病毒克隆载体使用的反向遗传系统。该系统代表了一种用于冠状病毒反向遗传操作的通用方法，最初是为产生代表I组冠状病毒的重组人冠状病毒229E而描述的。随后，相同的方法已被用于产生重组禽传染性支气管炎冠状病毒，并且最近还用于产生分别代表III组和II组冠状病毒的重组小鼠肝炎病毒。我们描述了痘苗病毒介导的同源重组如何在冠状病毒基因组cDNA于痘苗病毒中繁殖期间用于引入特定突变，以及如何分离重组冠状病毒。最后，我们描述了冠状病毒反向遗传系统现在如何已扩展到冠状病毒复制子RNA的产生。
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