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中国对虾过氧化氢酶抗氧化酶基因的cDNA克隆、特性分析及表达分析

cDNA cloning, characterization and expression analysis of the antioxidant enzyme gene, catalase, of Chinese shrimp Fenneropenaeus chinensis.

作者信息

Zhang Qingli, Li Fuhua, Zhang Xiaojun, Dong Bo, Zhang Jiquan, Xie Yusu, Xiang Jianhai

机构信息

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, PR China.

出版信息

Fish Shellfish Immunol. 2008 May;24(5):584-91. doi: 10.1016/j.fsi.2008.01.008. Epub 2008 Feb 1.

DOI:10.1016/j.fsi.2008.01.008
PMID:18353680
Abstract

Catalase is an important antioxidant protein that protects organisms against various oxidative stresses by eliminating hydrogen peroxide. The full-length catalase cDNA of Chinese shrimp Fenneropenaeus chinensis was cloned from the hepatopancreas using degenerate primers by the method of 3' and 5' rapid amplification of cDNA ends PCR. The cDNA sequence consists of 1892 bp with a 1560 bp open reading frame, encoding 520 amino acids with high identity to invertebrate, vertebrate and even bacterial catalases. The sequence includes the catalytic residues His71, Asn144, and Tyr354. The molecular mass of the predicted protein is 58824.04 Da with an estimated pI of 6.63. Sequence comparison showed that the deduced amino acid sequence of F. chinensis catalase shares 96%, 73%, 71% and 70% identity with that of Pacific white shrimp Litopenaeus vannamei, Abalone Haliotis discus hannai, Zhikong scallop Chlamys farreri and Human Homo sapiens, respectively. Catalase transcripts were detected in hepatopancreas, hemocytes, lymphoid organ, intestine, ovary, muscle and gill by real-time PCR. The variation of catalase mRNA transcripts in hemocytes and hepatopancreas was also quantified by real-time PCR and the result indicated that the catalase showed up-regulated expression trends in hemocytes at 14 h and in hepatopancreas at 37 h after injection with white spot syndrome virus (WSSV).

摘要

过氧化氢酶是一种重要的抗氧化蛋白,通过清除过氧化氢来保护生物体免受各种氧化应激。采用3'和5' cDNA末端快速扩增PCR方法,使用简并引物从中国对虾(凡纳滨对虾)的肝胰腺中克隆了其全长过氧化氢酶cDNA。该cDNA序列由1892 bp组成,开放阅读框为1560 bp,编码520个氨基酸,与无脊椎动物、脊椎动物甚至细菌的过氧化氢酶具有高度同源性。该序列包含催化残基His71、Asn144和Tyr354。预测蛋白的分子量为58824.04 Da,估计的等电点为6.63。序列比较表明,中国对虾过氧化氢酶推导的氨基酸序列与南美白对虾、皱纹盘鲍、栉孔扇贝和人类的氨基酸序列分别具有96%、73%、71%和70%的同源性。通过实时PCR在肝胰腺、血细胞、淋巴器官、肠道、卵巢、肌肉和鳃中检测到过氧化氢酶转录本。还通过实时PCR对血细胞和肝胰腺中过氧化氢酶mRNA转录本的变化进行了定量,结果表明,在注射白斑综合征病毒(WSSV)后14 h血细胞和37 h肝胰腺中过氧化氢酶呈现上调表达趋势。

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