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草鱼NADPH氧化酶2个调节亚基cDNA的克隆及表达特征
引用本文:吴涛,周剑光,张林.草鱼NADPH氧化酶2个调节亚基cDNA的克隆及表达特征[J].淡水渔业,2018(2):24-31.
作者姓名:吴涛  周剑光  张林
作者单位:1. 中国水产科学研究院长江水产研究所,武汉,430223;2. 武汉轻工业大学,武汉,430023
基金项目:中国水产科学研究院基本科研业务费
摘    要:为了解草鱼(Ctenopharyngodon idellus)NADPH多酶复合体的基因结构及其在机体的防御体系中的功能,利用RT-PCR结合RACE-PCR的方法,克隆草鱼NADPH氧化酶的2个调节亚基p40phox和p47phox的cDNA,对其编码的氨基酸序列进行了同源性比较和功能域分析,同时对这两个亚基在草鱼不同组织中的差异性表达进行分析。结果显示:p47phox亚基cDNA序列全长为1 589 bp,开放阅读框长度为1 233 bp,编码410个氨基酸;p40phox亚基cDNA序列全长为2 103 bp,开放阅读框为1 068 bp,编码355个氨基酸。这两个亚基编码的氨基酸序列与其他鱼类的同源性在68%~96%,具有其它鱼类类似的PX,SH3,PB1和PC功能域。组织差异性表达分析结果表明:2个调节亚基在草鱼胸腺、心脏、头肾、鳃、肠、肝、肾、脾和皮肤组织中均有表达,在不同组织中的表达强度略有差异,在心脏、胸腺中表达水平最高,在肝脏中表达水平较低,但是不同组织之间的表达水平差异不显著(P0.05)。

关 键 词:草鱼(Ctenopharyngodon  idellus)  NADPH氧化酶  分子克隆  组织表达  Ctenopharyngodon  idellus  NADPH  oxidase  molecular  cloning  tissue  expression

Cloning and expression characterizations of two regulatory subunits cDNA of NADPH oxidase from Ctenopharyngodon idellus
WU Tao,ZHOU Jian-guang,ZHANG Lin.Cloning and expression characterizations of two regulatory subunits cDNA of NADPH oxidase from Ctenopharyngodon idellus[J].Freshwater Fisheries,2018(2):24-31.
Authors:WU Tao  ZHOU Jian-guang  ZHANG Lin
Abstract:In order to study the genetic structure and function of the NADPH oxidase complexes of Ctenopharyngodon idel-lus, the full - length cDNAs of two regulatory subunits of NADPH oxidase, p40phox and p47phox were cloned from C.idellus by RT-PCR and RACE-PCR.The alignment of the peptide encoded by the two genes and function domains were analyzed, meanwhile the expression level of the two genes in different tissues were measured by Real time PCR.The results showed that the full length of p47phox cDNA was 1 589 bp, containing a 1 233 bp open reading frame that encoded a 410 amino acid protein.The full length of p40phox cDNA was 2 103 bp, containing a 1 068 bp open reading frame that encoded a 355 amino acid protein.Homology of the amino acid sequences encoded by the two genes to those of other fishes were from 68% to 96%, and they had similar functional domains of PX, SH3, PB1 and PC in other fishes.The results of expression level of the two genes in different tissues of C.idellus showed that the two subunits could be detected in thymus, heart, head-kidney, gill, intestines, liver, kidney, spleen and skin, their expression level were different in different tissues, with the highest level in heart and thymus, and with the lower level in liver, but there was no significant differ-ence between these tissues.
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