Molecular cloning and sequence analysis of the Danio rerio catalase gene
Molecular cloning and sequence analysis of the Danio rerio catalase gene
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DOI：
10.1016/s0305-0491(00)00285-6
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发表时间：
2000-12-01
影响因子：
2.2
通讯作者：
Grundy, MA
中科院分区：
文献类型：
--
作者：
Gerhard, GS;Kauffman, EJ;Grundy, MA
过氧化氢酶是一种抗氧化酶，通过过氧化氢的代谢在保护机体免受氧化应激中发挥核心作用。过氧化氢酶已经在植物、细菌和哺乳动物中得到了很好的研究，但在其他脊椎动物物种中的研究很少。我们克隆了斑马鱼过氧化氢酶cDNA的完整编码区，并通过逆转录聚合酶链反应和蛋白质印迹分析表达。推导的氨基酸序列预测了一个526个氨基酸的蛋白质，其一级DNA和氨基酸序列在脊椎动物物种中高度保守。过氧化氢酶复合物中的主要蛋白质-血红素接触点也很保守，尽管与主要底物通道的第二和第三水平相关的几个氨基酸不保守，这表明底物进入或特异性存在潜在差异。cDNA的3'侧翼区在终止密码子附近含有一个二核苷酸重复序列，由一个近乎完美的多态性CA阵列组成。大鼠和小鼠过氧化氢酶基因还在3'非翻译区中含有CA重复序列，其与相邻的5'茎环结构一起沿着，先前已显示为mRNA蛋白结合的位点（Clerch，1995，Arch.Biochem.Biophys.2004，2005）。317（1995）267-274）。一个茎环结构也预测邻近的斑马鱼CA重复，表明过氧化氢酶基因调控中的类似作用。(C)2000 Elsevier Science Inc. All rights reserved.
Catalase is an antioxidant enzyme that plays a central role in the protection against oxidative stress through the metabolism of hydrogen peroxide. Catalase has been well studied in plants, bacteria, and mammals, but little work has been done in other vertebrate species. We have cloned the zebrafish (Danio rerio) catalase cDNA containing the complete coding region and analyzed expression by both reverse transcription polymerase chain reaction and western blot. The deduced amino acid sequence predicts a protein of 526 amino acids with both the primary DNA and amino acid sequences highly conserved among vertebrate species. The major protein-heme contact points in the catalase enzyme complex are also well conserved, although several amino acids associated with the second and third levels of the major substrate channel are not, suggesting potential differences in substrate access or specificity. The 3' flanking region of the cDNA contains a dinucleotide repeat near the termination codon consisting of a near perfect CA array that is polymorphic. The rat and mouse catalase genes also contain a CA repeat sequence in the 3' untranslated region, which, along with an adjacent 5' stem-loop structure, has previously been shown to be a site for mRNA protein binding (Clerch, 1995, Arch. Biochem. Biophys. 317 (1995) 267-274). A stem-loop structure is also predicted adjacent to the zebrafish CA repeat, suggesting a similar role in catalase gene regulation. (C) 2000 Elsevier Science Inc. All rights reserved.