Identification of putative downstream genes of Oct-4 by suppression-subtractive hybridization
Identification of putative downstream genes of Oct-4 by suppression-subtractive hybridization
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DOI：
10.1006/bbrc.2001.4636
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发表时间：
2001-04-06
影响因子：
3.1
通讯作者：
Yao, Z
中科院分区：
文献类型：
--
作者：
Du, ZW;Cong, HC;Yao, Z
为了了解Oct-4是如何维持ToTI/多能性的，我们首先构建了在胚胎干细胞中差异表达Oct-4的细胞模型，然后利用抑制-消减杂交(SSH)方法鉴定了Oct-4的下游基因。在我们筛选的384个克隆中，40个克隆经菌落杂交鉴定为差异表达基因，13个克隆经Northern印迹分析证实为Oct-4的下游基因。测序发现12个不同的基因，8个已知基因(Oct-4、Rex-1、Sox-2、肌酸激酶B、makorin 1、Importinβ、组蛋白H2A.Z、核糖体蛋白S7)和4个新基因。除Oct-4和Rex-1外，其余基因均未受Oct-4调控，这些结果表明SSH为鉴定转录因子下游基因提供了一种非常有效的手段。一些已识别的已知基因可能为了解Oct-4在干细胞中的功能提供新的见解。(C)2001年学术出版社。
As a step toward understanding how toti/pluripotence is maintained by Oct-4, we have first constructed a cell model with differentially expressed Oct-4 in embryonic stem cells, and then used suppression-subtractive hybridization (SSH) method to identify the downstream genes of Oct-4. Among the 384 clones we screened, 40 clones were detected as differentially expressed genes with colony hybridization, and 13 clones were confirmed as the putative downstream genes of Oct-4 by Northern blot analysis. Sequencing showed 12 different genes, 8 known genes (Oct-4, Rex-1, Sox-2, Creatine kinase B, Makorin 1, Importin beta, Histone H2A.Z, Ribosomal protein S7) and 4 new genes. Except Oct-4 and Rex-1, the other genes have not been reported to be regulated by Oct-4, These results showed that SSH provides a very efficient means to identify the downstream genes of transcription factor. Some known genes identified may provide new insight of the function of Oct-4 in stem cells. (C) 2001 Academic Press.