ISOLATION OF ORC6, A COMPONENT OF THE YEAST ORIGIN RECOGNITION COMPLEX BY A ONE-HYBRID SYSTEM
ISOLATION OF ORC6, A COMPONENT OF THE YEAST ORIGIN RECOGNITION COMPLEX BY A ONE-HYBRID SYSTEM
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DOI：
10.1126/science.8266075
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发表时间：
1993-12-17
期刊：
影响因子：
56.9
通讯作者：
HERSKOWITZ, I
中科院分区：
文献类型：
--
作者：
LI, JJ;HERSKOWITZ, I
这里描述了一种方法，以确定基因编码蛋白质识别特定的DNA序列。筛选带有转录激活域标记的随机蛋白质片段库，以寻找能够激活包含其启动子序列的报告基因的蛋白质。这一策略被用来鉴定一种在体内与酵母起源的DNA复制相互作用的基本蛋白质。将预测的氨基酸序列与酵母起源识别复合体（ORC） 50千道尔顿亚基的肽序列进行比对，证实了分离的基因ORC6编码该亚基。这些观察结果提供了ORC识别酵母菌体内复制起源的证据。
Here a method is described to identify genes encoding proteins that recognize a specific DNA sequence. A bank of random protein segments tagged with a transcriptional activation domain is screened for proteins that can activate a reporter gene containing the sequence in its promoter. This strategy was used to identify an essential protein that interacts in vivo with the yeast origin of DNA replication. Matches between its predicted amino acid sequence and peptide sequence obtained from the 50-kilodalton subunit of the yeast origin recognition complex (ORC) established that the gene isolated here, ORC6, encodes this subunit. These observations provide evidence that ORC recognizes yeast replication origins in vivo.