Cu^(2+)胁迫下花斑裸鲤IL-8基因的组织表达分析

The Tissue Expression Analysis of the GeIL-8 Gene and the Response to Cu^(2+) Stress in Gymnocypris Eckloni

采用RT-PCR及cDNA末端快速扩增技术克隆了花斑裸鲤IL-8基因的全长及生物信息学特征,用Real-time PCR技术分析GeIL-8mRNA在花斑裸鲤各个组织中的表达分布,研究黄河上游水域水质污染的生物标记物,为水环境污染的早期预警以及花斑裸鲤的保护提供参考。结果表明:花斑裸鲤GeIL-8序列共582bp,由45bp的5'-UTR、240bp的3'-UTR和297bp的编码区构成,编码98个氨基酸;GeIL-8基因核苷酸序列与鲫鱼的同源性最高(87%)。Real-time PCR结果表明,正常条件下花斑裸鲤GeIL-8肝组织中的表达量最高,其次是鳃,在肾和脑组织中表达量较低;在Cu^(2+)胁迫下,花斑裸鲤肝脏、肾脏、脑和鳃组织的GeIL-8在mRNA水平的表达量总体均表现为先升后降的规律性应答,说明Cu^(2+)能够诱导GeIL-8的表达,也表明GeIL-8是潜在的反映水环境质量的生物标志物。研究结果提示,IL-8可作为黄河上游水域水质污染的生物标记物。

Copper is a necessary trace element in fish,acting as a cofactor for enzymes to participate in many physiological and biochemical processes in fish.But Cu^（2＋）is also a common heavy metal pollutant.When its content is too high,it will harm the neurophysiological function of fish,the structure of the olfactory organ and the endocrine.The paper described the cloning of the Gymnocypris eckloni interleukin-8 gene and analysed analyzed its characteristics and biological information,through which IL-8 response model in the liver and kidney,brain and gill tissues in different tissues under Cu^（2＋）stress was obtained.Rapid amplification of cDNA end g.eckloni and real-time PCR were used to clone the full-length GeIL-8 gene,and the mRNA expression and distribution of GeIL-8 in normal physiological conditions and under0.01 mg/L Cu^（2＋）stress in various tissues of Gymnocypris eckloni were analysed analyzed.The results showed that the GeIL-8 sequence was 582 bp,consisting of 45 bp 5＇-UTR,240 bp 3＇-UTR and 297 bp encoding region,encoding 98 amino acids;GeIL-8 gene nucleotide sequence shared homology with crucian carp（87%）.Real-time PCR showed that the expression of GeIL-8 in liver was the highest under normal conditions,followed by the gill,kidney and brain;under Cu^（2＋）stress,the mRNA expression of GeIL-8 in liver,kidney,brain and gill all showed a trend of first rising and then falling.It indicates that Cu^（2＋）can induce the expression of GeIL-8,also shows that GeIL-8 is the biological markers of water environment quality.