Human ORFeome version 1.1: A platform for reverse proteomics
Human ORFeome version 1.1: A platform for reverse proteomics
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DOI：
10.1101/gr.2973604
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发表时间：
2004-10-01
期刊：
影响因子：
7
通讯作者：
Vidal, M
中科院分区：
文献类型：
--
作者：
Rual, JF;Hirozane-Kishikawa, T;Vidal, M
系统生物学的出现需要克隆几乎一整套编码蛋白质的开放阅读框架(ORF)或ORFeome，以便对相应的蛋白质组进行功能研究。在这里，我们描述了使用一种新改进的Gateway重组克隆方法来生成第一个版本的人ORFeome。以哺乳动物基因库(MGC)资源为起点，我们报道了8076个人类ORF的成功克隆，代表至少7263个人类基因，作为小型的PCR扩增产物池。这些被组装到人类ORFeome版本1.1(hORFeome v1.1)集合中。在评估了该版本的整体质量之后，我们描述了hORFeome V1.1在蛋白质组规模的两个不同表达系统中的异源蛋白表达的使用。HORFeome V1.1代表了在各种环境中克隆大量人类ORF的核心资源，用于许多类型的功能蛋白质组学，并将作为后续人类ORFeome改进版本的基础。
The advent of systems biology necessitates the cloning of nearly entire sets of protein-encoding open reading frames (ORFs), or ORFeomes, to allow functional studies of the corresponding proteomes. Here, we describe the generation of a first version of the human ORFeome using a newly improved Gateway recombinational cloning approach. Using the Mammalian Gene Collection (MGC) resource as a starting point, we report the successful cloning of 8076 human ORFs, representing at least 7263 human genes, as mini-pools of PCR-amplified products. These were assembled into the human ORFeome version 1.1 (hORFeome v1.1) collection. After assessing the overall quality of this version, we describe the use of hORFeome v1.1 for heterologous protein expression in two different expression systems at proteome scale. The hORFeome v1.1 represents a central resource for the cloning of large sets of human ORFs in various settings for functional proteomics of many types, and will serve as the foundation for subsequent improved versions of the human ORFeome.