Cloning and characterization of the rabbit POU5F1 gene.

Jian J Shi, Dong H Cai, Xue J Chen, Hui Z Sheng
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引用次数: 9

Abstract

The product of the POUSF1 gene, Oct4, plays an important role both in embryonic development and in the self-renewal and differentiation of totipotent cells. To understand the function of Oct4 in rabbit ES cells, we cloned and sequenced the rabbit POU5F1 gene, as well as the cDNA encoded by the gene. The Oct4 cDNA contains a 1083 bp ORF encoding a 360 aa protein and a 241 bp 3' UTR sequence. Oct4 mRNA was expressed at a high level in rabbit ES cells and was barely detectable in the adult spleen, kidney, brain and muscle tissues. The POU5F1 gene is approximately 6 kb in length and includes five exons and four introns. Gene organization is similar to that of the mouse, human and bovine orthologs. Sequencing of the gene revealed an 82% (mouse), 90% (human) and 89% (bovine) overall identity at the protein level. The rabbit POUSF1 gene was mapped to chromosome 12q1.1 by PCR amplification of DNA from two putative POU5F1-containing BAC clones, which were previously mapped to chromosome 12q1.1. The cloning of the rabbit POU5F1 gene will facilitate studies on its roles in rabbit embryogenesis and ES cells.

家兔POU5F1基因的克隆与鉴定。
POUSF1基因的产物Oct4在胚胎发育和全能细胞的自我更新和分化中都起着重要作用。为了了解Oct4在兔胚胎干细胞中的功能,我们克隆并测序了兔POU5F1基因,以及该基因编码的cDNA。Oct4 cDNA包含一个1083 bp的ORF,编码一个360 aa蛋白和一个241 bp的3' UTR序列。Oct4 mRNA在兔胚胎干细胞中高水平表达,在成人脾、肾、脑和肌肉组织中几乎检测不到。POU5F1基因全长约6kb,包括5个外显子和4个内含子。基因组织与小鼠、人类和牛的同源物相似。测序结果显示,该基因在蛋白质水平上的总体一致性为82%(小鼠)、90%(人类)和89%(牛)。兔POUSF1基因被定位到染色体12q1.1上,通过PCR扩增两个假定含有pou5f1的BAC克隆的DNA。兔POU5F1基因的克隆将有助于研究其在兔胚胎发生和胚胎干细胞中的作用。
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