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在中国北京高碑店湖受污染物和热暴露的金鱼(Carassius auratus)中差异表达基因的鉴定。

Identification of differentially expressed genes from contaminant and thermal exposed goldfish Carassius auratus in Gaobeidian Lake in Beijing, China.

作者信息

Wang Jianshe, Wei Yanhong, Li Xuemei, Xu Muqi, Dai Jiayin

机构信息

Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, P.R. China.

出版信息

Ecotoxicology. 2007 Oct;16(7):525-32. doi: 10.1007/s10646-007-0159-8. Epub 2007 Jul 11.

Abstract

Gaobeidian Lake, located in Beijing, China, derives its water mainly from the effluent of the Gaobeidian Wastewater Treatment Plant, which is moderately polluted. Additionally, as this water is used as a coolant in the nearby thermal power plant, the water of this lake has an elevated temperature. To screen differential gene expression in Gaobeidian Lake, suppressive subtractive hybridization (SSH) methodology was performed on RNA in goldfish Carassius auratus hepatic tissues from the lake, using Huairou Reservoir as reference site. A total of 768 candidate clones were selected to perform differential screening. Of these, 264 clones were differentially expressed between the two sites, 124 of which were then subjected to DNA sequencing. Consequently, 36 different genes with known functions were obtained, and some of these differential genes were further confirmed by semiquantitative RT-PCR experiments. Many genes related to detoxification, stress and immune response, and metabolism, such as glutathione S-transferase (GST), cytochrome P450, family 2, subfamily b, polypeptide 10 (CYP2B10), CYP2X10, alpha-1-antitrypsin, and apolipoprotein A-I (Apo-AI), had higher expression levels in goldfish hepatic tissue from Gaobeidian Lake than those from the reference site. A set of nine genes with known functions were downregulated in Gaobeidian Lake compared to the reference site. The results provided evidence that organisms inhabiting Gaobeidian Lake were suffering a complex stress process and showing metabolism changes and disturbance of homeostasis.

摘要

高碑店湖位于中国北京,其水源主要来自高碑店污水处理厂的出水,水质为中度污染。此外,由于该湖水被附近的热电厂用作冷却用水,湖水温度有所升高。为筛选高碑店湖中的差异基因表达,以怀柔水库为对照点,采用抑制性消减杂交(SSH)技术对采自该湖的金鱼肝脏组织RNA进行分析。共挑选出768个候选克隆进行差异筛选。其中,264个克隆在两个采样点间存在差异表达,从中选取124个进行DNA测序。最终获得36个功能已知的差异基因,部分差异基因通过半定量RT-PCR实验进一步得到验证。许多与解毒、应激和免疫反应以及代谢相关的基因,如谷胱甘肽S-转移酶(GST)、细胞色素P450 2B亚家族10型(CYP2B10)、CYP2X10、α-1-抗胰蛋白酶和载脂蛋白A-I(Apo-AI),在来自高碑店湖的金鱼肝脏组织中的表达水平高于对照点。与对照点相比,高碑店湖中有9个功能已知的基因表达下调。结果表明,栖息于高碑店湖的生物正经历复杂的应激过程,代谢发生变化,体内稳态受到干扰。

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