槟榔Catalase基因的克隆及亚细胞定位

过氧化氢酶（Catalase）广泛存在于生物体内,主要功能是清除植物细胞内光呼吸、线粒体电子传递及脂肪β-氧化等过程中产生的H₂O₂,从而保护细胞免于过氧化氢酶的毒害。为了解槟榔过氧化氢酶基因的相关信息,利用植物总RNA提取试剂盒提取槟榔叶片总RNA,通过同源克隆和RACE技术获得槟榔全长cDNA,并对其进行生物信息学分析及亚细胞定位。结果表明：获得ArCAT1（MN692600）、ArCAT2（MN692601）、ArCAT3（MN692602）3个同源基因,其序列全长均为1476 bp,编码492个氨基酸。遗传进化分析显示,ArCATS与同为棕榈科的油棕（Elaeis guineensis）、海枣（Phoenix dactylifera）等植物的过氧化氢酶氨基酸序列具有较高的相似性,其中与油棕的亲缘关系最近。亚细胞定位结果表明,ArCAT1蛋白定位于过氧化物酶体中,而ArCAT2和ArCAT3既定位于过氧化物酶体又定位于细胞核中。这为进一步研究Catalase基因在槟榔中的生物学功能奠定基础。

Cloning and Subcellular Localization of Catalase Gene in Areca catechu L.

The main function of catalase, widely existed in organisms, is to remove H₂O₂ produced in the process of photorespiration, mitochondrial electron transmission and fat betao-oxidation in plant cells to protect cells from the toxicity of H₂O₂. In order to understand the relevant information of areca catalase gene, RNA was extracted from areca leaves, the full length of areca cDNA was obtained through the homologous cloning and RACE technology, and bioinformatics analysis and subcellular localization were performed. The results showed that the total length of ArCAT1 (MN692600), ArCAT2 (MN692601) and ArCAT3 (MN692602) gene sequence was 1476 bp, which could encode 492 amino acids. Genetic evolutionary analysis showed that ArCATS was closely related to Elaeis guineensis and Phoenix dactylifera. The results of subcellular localization showed that ArCAT1 protein was located in the peroxidosome, ArCAT2 and ArCAT3 were located in the peroxidosome and nucleus. This would lay foundation for further research on the biological function of the gene.