青岛文昌鱼过氧化氢酶基因的克隆及生物信息学分析

过氧化氢酶(catalase,CAT)是一类广泛存在于动物、植物和微生物体内的末端氧化酶,是生物体内抗氧化酶体系的重要成员,因此本研究旨在克隆文昌鱼CAT基因并对其进行生物信息学分析。本研究以青岛文昌鱼(Branchiostoma belcheri tsingtauense)为材料,用RACE技术首次克隆了其CAT全长cDNA的基因序列,命名为AmphiCAT(GenBank登录号:KU058636);该序列总长为2640 bp,开放阅读框(open reading frame,ORF)为1533 bp,编码510个氨基酸,含有一个长为19个氨基酸序列的潜在活性位点和一个长为9个氨基酸序列的血红素配体信号,总分子量在线预测约为57.85 kD;经生物软件分析确定该蛋白质无信号肽序列,预测该蛋白质为非分泌性蛋白,属于单功能CAT的clade3分支;该基因的分子进化树表明青岛文昌鱼CAT同软体动物的亲缘关系较近。

Cloning and Bioinformatics Analysis of Catalase Gene from Branchiostoma belcheri tsingtauense

Catalase is a kind of terminaloxidase that widely exists in animals,plants and microorganisms,which is an important member in antioxidant enzyme system.This study aims to explore cloning catalase gene of amphioxus and its bioinformatics analysis.In this study,the full-length cDNA of catalase(CAT)gene named AmphiCAT(GenBank accession No.KU058636)was cloned and characterized using rapid amplification of cDNA ends(RACE)from Branchiostoma belcheri tsingtauense.The full-length cDNA of CAT was 2640 bp,including1533 bp open reading frame(ORF)encoding 510 amino acids,containing a potential active site with a length of19 amino acid sequences and a heme ligand signal with a length of 9 amino acid sequences.The total molecular weight was estimated online to be about 57.85 kD.The protein was determined that it was no signal peptide sequence by biological software analysis,and it was predicted that the protein was non-secretory protein,belonging to clade 3 branch of monofunctional catalase.Phylogenetic analysis suggested that amphioxus had close relatives with mollusks.