INSERM U851, Lyon, IFR128-BioSciences, Université Lyon 1, Université de Lyon, Lyon, France.
Nucleic Acids Res. 2010 Jan;38(Database issue):D371-8. doi: 10.1093/nar/gkp1000. Epub 2009 Dec 8.
Large collections of protein-encoding open reading frames (ORFs) established in a versatile recombination-based cloning system have been instrumental to study protein functions in high-throughput assays. Such 'ORFeome' resources have been developed for several organisms but in virology, plasmid collections covering a significant fraction of the virosphere are still needed. In this perspective, we present ViralORFeome 1.0 (http://www.viralorfeome.com), an open-access database and management system that provides an integrated set of bioinformatic tools to clone viral ORFs in the Gateway(R) system. ViralORFeome provides a convenient interface to navigate through virus genome sequences, to design ORF-specific cloning primers, to validate the sequence of generated constructs and to browse established collections of virus ORFs. Most importantly, ViralORFeome has been designed to manage all possible variants or mutants of a given ORF so that the cloning procedure can be applied to any emerging virus strain. A subset of plasmid constructs generated with ViralORFeome platform has been tested with success for heterologous protein expression in different expression systems at proteome scale. ViralORFeome should provide our community with a framework to establish a large collection of virus ORF clones, an instrumental resource to determine functions, activities and binding partners of viral proteins.
大量基于重组的通用克隆系统建立的蛋白质编码开放阅读框 (ORF) 集合,对于在高通量测定中研究蛋白质功能非常重要。已经为几种生物体开发了这样的“ORFeome”资源,但在病毒学中,仍然需要涵盖病毒圈很大一部分的质粒集合。在这方面,我们展示了 ViralORFeome 1.0(http://www.viralorfeome.com),这是一个开放访问的数据库和管理系统,提供了一套集成的生物信息学工具,用于在 Gateway(R)系统中克隆病毒 ORF。ViralORFeome 提供了一个方便的界面,可以浏览病毒基因组序列,设计 ORF 特异性的克隆引物,验证生成的构建体的序列,并浏览已建立的病毒 ORF 集合。最重要的是,ViralORFeome 旨在管理给定 ORF 的所有可能变体或突变体,以便克隆程序可以应用于任何新出现的病毒株。已经使用 ViralORFeome 平台生成的质粒构建体的子集在不同的表达系统中进行了异源蛋白质表达的大规模蛋白质组测试,取得了成功。ViralORFeome 应该为我们的社区提供建立大量病毒 ORF 克隆集合的框架,这是确定病毒蛋白功能、活性和结合伙伴的重要资源。