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斑马鱼(Danio rerio)锌指转录因子MTF-1基因的分子克隆与发育表达

Molecular cloning and developmental expression of zinc finger transcription factor MTF-1 gene in zebrafish, Danio rerio.

作者信息

Chen Wen-Ya, John Joseph Abraham Christopher, Lin Chen-Hui, Chang Chi-Yao

机构信息

Institute of Zoology, Academia Sinica, Taiwan, NanKang, Taipei, Republic of China.

出版信息

Biochem Biophys Res Commun. 2002 Mar 8;291(4):798-805. doi: 10.1006/bbrc.2002.6517.

Abstract

Metal-responsive transcription factor, MTF-1 is a zinc finger protein, shown to be essential for embryonic development. Homozygous knockout mouse embryos for MTF-1 die in utero at day 14 of gestation, due to liver decay. In the present study, we report the complete nucleotide sequence of cDNA encoding zebrafish MTF-1 and the amino acid sequence similarity with that of mouse, human, fish and Drosophila. The size of the zebrafish MTF-1 cDNA is 3,379 bp and the coding region (1,779 bp) encodes a polypeptide of 593 amino acids. The putative zinc finger and transactivation domains comprised by zebrafish MTF-1 were also determined. The zebrafish MTF-1 shows high identity of 97, 93, 93 and 67% in the DNA binding zinc finger domain and 51, 44, 48 and 20% overall identity with fugu, human, mouse and Drosophila, respectively. RT-PCR results show the maternal expression of MTF-1 transcripts. The pattern of MTF-1 gene expression during embryonic and early larval development was studied by whole-mount in situ hybridization using DIG-labeled anti-sense RNA probe. Stronger and ubiquitous expression was observed during the embryonic stages whereas, specific expression, especially in the neural parts, was observed throughout the stages studied after hatching.

摘要

金属反应转录因子MTF-1是一种锌指蛋白,已证明对胚胎发育至关重要。MTF-1基因纯合敲除的小鼠胚胎在妊娠第14天死于子宫内,原因是肝脏衰退。在本研究中,我们报告了编码斑马鱼MTF-1的cDNA的完整核苷酸序列以及与小鼠、人类、鱼类和果蝇的氨基酸序列相似性。斑马鱼MTF-1 cDNA的大小为3379 bp,编码区(1779 bp)编码一个由593个氨基酸组成的多肽。还确定了斑马鱼MTF-1所含的推定锌指和反式激活结构域。斑马鱼MTF-1在DNA结合锌指结构域中与河豚、人类、小鼠和果蝇的同源性分别为97%、93%、93%和67%,总体同源性分别为51%、44%、48%和20%。RT-PCR结果显示MTF-1转录本的母源性表达。使用地高辛标记的反义RNA探针通过整体原位杂交研究了MTF-1基因在胚胎和幼体早期发育过程中的表达模式。在胚胎阶段观察到更强且普遍的表达,而在孵化后的整个研究阶段观察到特异性表达,尤其是在神经部位。

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