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Suppression subtractive hybridization library prepared from the copepod Calanus finmarchicus exposed to a sublethal mixture of environmental stressors

机译:由暴露于环境致死因子亚致死混合物中的pe足类Calaus finmarchicus制备的抑制性消减杂交文库

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摘要

A library of expressed sequence tags (ESTs) was constructed by the use of suppression subtractive hybridization polymerase chain reaction (SSH PCR) technique from the marine copepod Calanus finmarchicus. Samples used were from controls (seawater, 10 °C) and exposed (sublethal mixture) individuals. The sublethal exposure regime consisted of a mixture of mono ethanol amine (MEA), water-soluble fractions of oil (WSFs), copper (Cu) and elevated temperature (17 °C). The resulting 189 unique ESTs consisted of 127 putatively up-regulated genes and 54 putatively down-regulated genes. Annotation analyses revealed altered expression of a wide variety of genes, among these putative heat shock protein 90 (HSP-90), antioxidants (thioredoxin reductase, glutathione peroxidase) and cytochrome P450 enzymes. In addition, sequences showing high similarity to enzymes involved in fatty acid metabolism, energy metabolism and amine handling were found further confirming the effects of the exposure. The annotated sequences are discussed in relation to the present exposure as well as known physiological mechanisms known in C. finmarchicus and related copepod species. The sequenced ESTs from our C. finmarchicus library will provide an excellent tool for future studies on this species, both from a toxicogenomic and systems biology point of view.
机译:利用抑制suppression消减杂交聚合酶链反应(SSH PCR)技术,从海洋pe足类Calaus finmarchicus构建了表达序列标签(EST)文库。使用的样品来自对照(海水,10°C)和暴露的(亚致死混合物)个体。亚致死暴露方案由单乙醇胺(MEA),油的水溶性馏分(WSFs),铜(Cu)和高温(17°C)的混合物组成。产生的189个独特EST包含127个可能被上调的基因和5​​4个可能被下调的基因。注释分析显示,在这些假定的热休克蛋白90(HSP-90),抗氧化剂(硫氧还蛋白还原酶,谷胱甘肽过氧化物酶)和细胞色素P450酶中,多种基因的表达发生了变化。另外,发现与脂肪酸代谢,能量代谢和胺处理中涉及的酶显示高度相似性的序列进一步证实了暴露的影响。讨论了与当前暴露有关的注释序列,以及在翅翅隐孢菌和相关co足类物种中已知的已知生理机制。从毒物基因组学和系统生物学的角度出发,我们的芬克C. finmarchicus文库中测序的ESTs将为今后对该物种的研究提供一个极好的工具。

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