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鲤鱼毛状/分裂增强相关基因her6的克隆、特征分析及启动子分析

Cloning, characterization and promoter analysis of common carp hairy/Enhancer-of-split-related gene, her6.

作者信息

Liu Jing, Sun Yong-Hua, Wang Na, Wang Ya-Ping, Zhu Zuo-Yan

机构信息

State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.

出版信息

J Genet. 2006 Dec;85(3):171-8. doi: 10.1007/BF02935327.

Abstract

Some members of hairy/Enhancer-of-split-related gene (HES) family have important effects on axial mesoderm segmentation and the establishment and maintenance of the somite fringe. In fishes, the her6 gene, a member of the HES family, is the homologue of hes1 in mammals and chicken. In this study, the her6 gene and its full-length cDNA from the common carp (Cyprinus carpio) were isolated and characterized. The genomic sequence of common carp her6 is approximately 1.7 kb, with four exons and three introns, and the full-length cDNA of 1314 bp encodes a putative polypeptide of 271 amino acids. To analyse the promoter sequence of common carp her6, sequences of various lengths upstream from the transcription initiation site of her6 were fused to enhanced green fluorescent protein gene (eGFP) and introduced into zebrafish embryos by microinjection to generate transgenic embryos. Our results show that the upstream sequence of 500 bp can direct highly efficient and tissue-specific expression of eGFP in zebrafish embryos, whereas a fragment of 200 bp containing the TATA box and a partial suppressor of hairless paired site sequence (SPS) is not sufficient to drive eGFP expression in zebrafish embryos.

摘要

毛状/分裂增强子相关基因(HES)家族的一些成员对轴中胚层分割以及体节边缘的建立和维持具有重要作用。在鱼类中,HES家族成员her6基因是哺乳动物和鸡中hes1的同源物。在本研究中,分离并鉴定了鲤鱼(Cyprinus carpio)的her6基因及其全长cDNA。鲤鱼her6的基因组序列约为1.7 kb,有四个外显子和三个内含子,1314 bp的全长cDNA编码一个由271个氨基酸组成的假定多肽。为了分析鲤鱼her6的启动子序列,将her6转录起始位点上游不同长度的序列与增强型绿色荧光蛋白基因(eGFP)融合,并通过显微注射导入斑马鱼胚胎以产生转基因胚胎。我们的结果表明,500 bp的上游序列可在斑马鱼胚胎中指导eGFP的高效和组织特异性表达,而包含TATA盒和部分无毛配对位点序列(SPS)抑制子的200 bp片段不足以驱动斑马鱼胚胎中eGFP的表达。

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