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参与保护家蚕对氧化应激的过氧化物酶 4 的分子克隆与特性分析。

Molecular cloning and characterization of peroxiredoxin 4 involved in protection against oxidative stress in the silkworm Bombyx mori.

机构信息

College of Life Sciences, Chongqing University, China.

出版信息

Insect Mol Biol. 2012 Dec;21(6):581-92. doi: 10.1111/j.1365-2583.2012.01161.x. Epub 2012 Sep 5.

Abstract

Peroxiredoxins (Prxs) are a ubiquitous family of proteins that play important roles in insects in protection against oxidative stress through the detoxification of cellular peroxides. Here, we describe the cloning and characterization of a Prx4 cDNA of the silkworm Bombyx mori (BmPrx4). The BmPrx4 gene has an open reading frame of 744 bp encoding 248 amino acids and a conserved motif, VCP, involved in its presumed redox functions. The heterologously expressed proteins of the gene in Escherichia coli showed antioxidant activity, removed hydrogen peroxide and protected DnA. Western blotting analysis showed the presence of BmPrx4 in the haemolymph, suggesting that the protein is secretable. Moreover, BmPrx4 was expressed at all developmental stages. The expression level of BmPrx4 was relatively low during the feeding stage but high at the wandering stage. BmPrx4 was induced by quercetin or temperature stress. Immunohistochemical analysis revealed that BmPrx4 is present in the brain, neurones and olfactory organ of the head in silkworms. Overall, our results indicate that the expression profile of BmPrx4 correlates well with protection from oxidative damage. Our data provide clues for the development of control technology for agricultural and forestry pests as the silkworm is a representative of lepidopteran pests.

摘要

过氧化物酶(Prxs)是一种普遍存在的蛋白质家族,在昆虫中通过细胞过氧化物的解毒来保护它们免受氧化应激的影响,从而发挥着重要作用。在这里,我们描述了家蚕(Bombyx mori)Prx4 cDNA 的克隆和特性。BmPrx4 基因的开放阅读框长 744bp,编码 248 个氨基酸,具有一个保守基序 VCP,该基序参与其假定的氧化还原功能。该基因在大肠杆菌中的异源表达蛋白具有抗氧化活性,可去除过氧化氢并保护 DNA。Western blot 分析表明 BmPrx4 存在于血液中,表明该蛋白是可分泌的。此外,BmPrx4 在所有发育阶段都有表达。BmPrx4 在取食阶段的表达水平相对较低,但在 wandering 阶段较高。BmPrx4 可被槲皮素或温度应激诱导。免疫组织化学分析表明 BmPrx4 存在于家蚕的大脑、神经元和头部嗅觉器官中。总的来说,我们的结果表明 BmPrx4 的表达谱与氧化损伤的保护密切相关。我们的数据为农业和林业害虫的控制技术的发展提供了线索,因为家蚕是鳞翅目害虫的代表。

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