ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombe
ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombe
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DOI：
10.1038/nbt1222
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发表时间：
2006-07-01
影响因子：
46.9
通讯作者：
Yoshida, Minoru
中科院分区：
文献类型：
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作者：
Matsuyama, Akihisa;Arai, Ritsuko;Yoshida, Minoru
克隆生物体的蛋白质编码开放阅读框架（ORF）或“ORFeome”的整个集合是将基因组与下游“组学”应用连接起来的一种手段。在这里，我们报告了裂殖酵母裂殖酵母的ORFeome克隆的基础上的蛋白质组规模的研究。利用基于重组的克隆系统，我们获得了4，910个ORF，其形式易于用于各种分析。首先，我们通过表达在3'末端标记的每个ORF来评估在裂殖酵母基因组计划中的ORF预测。接下来，我们通过用黄色荧光蛋白标记每个ORF，确定了4，431个蛋白质的定位，相当于90%的裂殖酵母蛋白质组。此外，使用来普霉素B，一种核输出蛋白Crm1的抑制剂，我们确定了285种蛋白质，其定位受Crm1调节。
Cloning of the entire set of an organism's protein-coding open reading frames (ORFs), or 'ORFeome', is a means of connecting the genome to downstream 'omics' applications. Here we report a proteome-scale study of the fission yeast Schizosaccharomyces pombe based on cloning of the ORFeome. Taking advantage of a recombination-based cloning system, we obtained 4,910 ORFs in a form that is readily usable in various analyses. First, we evaluated ORF prediction in the fission yeast genome project by expressing each ORF tagged at the 3' terminus. Next, we determined the localization of 4,431 proteins, corresponding to similar to 90% of the fission yeast proteome, by tagging each ORF with the yellow fluorescent protein. Furthermore, using leptomycin B, an inhibitor of the nuclear export protein Crm1, we identified 285 proteins whose localization is regulated by Crm1.