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对虾感染 WSSV 过程中过氧化物酶的分子克隆、表达分析及特性研究。

Molecular cloning, expression analysis and characterization of peroxiredoxin during WSSV infection in shrimp Fenneropenaeus indicus.

机构信息

Centre for Biotechnology, Anna University, Chennai 600 025, Tamil Nadu, India.

出版信息

J Invertebr Pathol. 2012 Jan;109(1):52-8. doi: 10.1016/j.jip.2011.09.006. Epub 2011 Sep 24.

DOI:10.1016/j.jip.2011.09.006
PMID:21964313
Abstract

Innate immunity is the first line of defense in shrimps against invading pathogenic microorganisms. Peroxiredoxins (PRX) are the family of antioxidant proteins that play a crucial role in reduction of oxidative stress in host during viral infection. Peroxiredoxin from Fenneropenaeus indicus was identified, cloned and expressed in prokaryotic expression system. The Fi-PRX protein consists of two conserved Cys residues and belongs to typical 2-cys PRX family. The phylogenetic analysis revealed a close evolutionary relatedness of Fi-PRX with the PRX from Drosophila melanogaster PRX1 and distant origin with PRX sequences from other shrimp isolates Fenneropenaeus chinensis, Litopenaeus vannamei and Penaeus japonicus. Fi-PRX transcripts are constitutively expressed in hemocytes and tissues (gills, heart and muscle) and down regulated during 12 h, 24 h and 48 h of WSSV challenged shrimps. Fi-PRX protein levels correlated well with the corresponding levels of Fi-PRX transcripts in hemocytes and tissues of WSSV challenged shrimps. Recombinant Fi-PRX reduces insulin only in the presence of DTT suggesting that the antioxidant function of the protein is thiol dependent. These findings suggest that antioxidant activity of Fi-PRX play a significant role in neutralization of excessive free radicals and ROS generated during viral invasion.

摘要

先天免疫是虾类抵御入侵的致病性微生物的第一道防线。过氧化物还原酶(PRX)是抗氧化蛋白家族,在病毒感染宿主时对减少氧化应激起着至关重要的作用。从印度对虾中鉴定、克隆并在原核表达系统中表达了过氧化物还原酶。Fi-PRX 蛋白由两个保守的 Cys 残基组成,属于典型的 2-cys PRX 家族。系统发育分析表明,Fi-PRX 与果蝇 PRX1 的 PRX 具有密切的进化关系,而与中国对虾、凡纳滨对虾和日本对虾等其他虾类分离株的 PRX 序列的起源较远。Fi-PRX 转录本在血细胞和组织(鳃、心脏和肌肉)中持续表达,并在 WSSV 感染虾的 12、24 和 48 小时时下调。Fi-PRX 蛋白水平与 WSSV 感染虾的血细胞和组织中相应的 Fi-PRX 转录本水平密切相关。重组 Fi-PRX 仅在存在 DTT 的情况下才会还原胰岛素,这表明该蛋白的抗氧化功能依赖于巯基。这些发现表明,Fi-PRX 的抗氧化活性在中和病毒入侵过程中产生的过量自由基和 ROS 方面发挥了重要作用。

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