Biochemical and genetic analysis of the yeast proteome with a movable ORF collection
Biochemical and genetic analysis of the yeast proteome with a movable ORF collection
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DOI：
10.1101/gad.1362105
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发表时间：
2005-12-01
影响因子：
10.5
通讯作者：
Grayhack, EJ
中科院分区：
文献类型：
--
作者：
Gelperin, DM;White, MA;Grayhack, EJ
蛋白质组的功能分析是基因组研究的重要组成部分。为了促进不同的蛋白质组学方法，构建了5854个酵母表达质粒的MORF（可移动ORF）文库，每个质粒在调控下表达序列验证的ORF作为C-末端ORF融合蛋白。对5573个MORFs的分析表明，几乎所有已验证的ORF都表达，表明48个ORF的真实性是可疑的，并暗示了特定的过程，包括细胞骨架组织和转录控制过度表达引起的生长抑制。糖基化蛋白的全球分析确定了109个新确认的N-连接和345个候选糖蛋白，几乎是已知酵母糖组的两倍。
Functional analysis of the proteome is an essential part of genomic research. To facilitate different proteomic approaches, a MORF (moveable ORF) library of 5854 yeast expression plasmids was constructed, each expressing a sequence-verified ORF as a C-terminal ORF fusion protein, under regulated control. Analysis of 5573 MORFs demonstrates that nearly all verified ORFs are expressed, suggests the authenticity of 48 ORFs characterized as dubious, and implicates specific processes including cytoskeletal organization and transcriptional control in growth inhibition caused by overexpression. Global analysis of glycosylated proteins identifies 109 new confirmed N-linked and 345 candidate glycoproteins, nearly doubling the known yeast glycome.