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在蜜蜂幼虫(中华蜜蜂)暴露于汞的情况下,对 sigma 类谷胱甘肽 S-转移酶基因进行了表征。

Characterization of a sigma class glutathione S-transferase gene in the larvae of the honeybee (Apis cerana cerana) on exposure to mercury.

机构信息

College of Life Sciences, Shandong Agricultural University, Taian, Shandong, 271018, PR China.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2012 Apr;161(4):356-64. doi: 10.1016/j.cbpb.2011.12.009. Epub 2012 Jan 8.

Abstract

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainly involved in detoxification of endogenous and xenobiotic compounds and oxidative stress resistance in insects. In this study, we identified a sigma class GST from Apis cerana cerana (AccGSTs4). The open reading frame of cDNA was 612 bp and encoded a 203 amino acid polypeptide, which exhibited the structural motif and domain organization characteristic of GST. Homology and evolutionary analysis indicated that the induced amino acid sequence of AccGSTs4 belonged to an insect sigma class group. Expression analysis indicated that AccGSTs4 was presented in all stages of development with high level in 4th instar larvae. Immunolocalization further revealed the distribution of AccGSTs4 in 4th instar larvae. RT-qPCR showed that the transcripts of AccGSTs4 from the larvae were upregulated under dietary HgCl(2). The GST activity under stress was higher than the controls fed on HgCl(2)-free diet. Disc diffusion assay provided evidence of recAccGSTs4 resistance to long-term exposure of HgCl(2) stress. Additionally, analysis of 5'-flanking region further clarified the probable expression patterns of AccGSTs4. Taken together, our findings indicate that the larvae AccGSTs4 may play a role in mercury stress response, and it will help to protect honeybees from heavy metals.

摘要

谷胱甘肽 S-转移酶(GSTs)是多功能酶,主要参与昆虫内源性和外源性化合物的解毒和氧化应激抗性。在本研究中,我们从中华蜜蜂(Apis cerana cerana)中鉴定出一种σ类 GST(AccGSTs4)。cDNA 的开放阅读框为 612bp,编码 203 个氨基酸的多肽,其表现出 GST 的结构基序和结构域组织特征。同源性和进化分析表明,AccGSTs4 的诱导氨基酸序列属于昆虫σ类群。表达分析表明,AccGSTs4 在所有发育阶段都有表达,在 4 龄幼虫中表达水平较高。免疫定位进一步揭示了 AccGSTs4 在 4 龄幼虫中的分布。RT-qPCR 显示,幼虫中 AccGSTs4 的转录本在饮食 HgCl2 存在下被上调。应激下的 GST 活性高于未摄入 HgCl2 的对照组。圆盘扩散试验为 recAccGSTs4 对长期 HgCl2 应激的抗性提供了证据。此外,对 5'侧翼区的分析进一步阐明了 AccGSTs4 的可能表达模式。总之,我们的研究结果表明,幼虫 AccGSTs4 可能在汞应激反应中发挥作用,并有助于保护蜜蜂免受重金属的侵害。

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