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金鱼(Carassius auratus)胰岛素样生长因子结合蛋白-2(IGFBP-2)cDNA的克隆、分子特征及表达分析

Cloning, molecular characterization and expression analysis of insulin-like growth factor binding protein-2 (IGFBP-2) cDNA in goldfish, Carassius auratus.

作者信息

Chen Wenbo, Li Weiguo, Zhang Zhen, Jiang Xiaoxue, Li Mengjie

机构信息

Department of Biology, Institute of Resources and Environment, Henan Polytechnic University, Jiaozuo, 454000, Henan, People's Republic of China,

出版信息

Fish Physiol Biochem. 2014 Dec;40(6):1669-81. doi: 10.1007/s10695-014-9958-z. Epub 2014 Jul 4.

DOI:10.1007/s10695-014-9958-z
PMID:24992902
Abstract

In the present study, a full-length cDNA encoding the insulin-like growth factor binding protein-2 (IGFBP-2) was cloned from the liver of goldfish (Carassius auratus) by rapid amplification of cDNA ends technique. The goldfish IGFBP-2 cDNA sequence was 1,513 bp long and had an open reading frame of 825 bp encoding a predicted polypeptide of 274 amino acid residues. Semi-quantitative RT-PCR results revealed that goldfish IGFBP-2 mRNA was expressed in all detected tissues. In liver, central nervous system and pituitary gland, goldfish IGFBP-2 expressed at high levels, followed by anterior intestine, middle intestine and kidney. In posterior intestine, ovary, skin, fat, spleen, muscle and gill, the goldfish IGFBP-2 expression levels were very low. Fasting and refeeding experiment showed that the mRNA expression of goldfish IGFBP-2 was up-regulated significantly in liver compared to the fed group and restored rapidly to normal level after refed. However, the mRNA expressions of IGFBP-2 in hypothalamus and pituitary of goldfish were insensitive to fasting. Furthermore, the mRNA expressions of IGFBP-2 in hypothalamus, pituitary and liver were varied in periprandial changes and significantly down-regulated at 2 and 4 h after meal. These results imply that the IGFBP-2 mRNA expression may be associated with anabolic and catabolic metabolism and regulated by metabolic factors in goldfish.

摘要

在本研究中,通过cDNA末端快速扩增技术从金鱼(Carassius auratus)肝脏中克隆了编码胰岛素样生长因子结合蛋白-2(IGFBP-2)的全长cDNA。金鱼IGFBP-2 cDNA序列长1513 bp,具有一个825 bp的开放阅读框,编码一个预测的含274个氨基酸残基的多肽。半定量RT-PCR结果显示,金鱼IGFBP-2 mRNA在所有检测组织中均有表达。在肝脏、中枢神经系统和垂体中,金鱼IGFBP-2高水平表达,其次是前肠、中肠和肾脏。在金鱼的后肠、卵巢、皮肤、脂肪、脾脏、肌肉和鳃中,IGFBP-2表达水平非常低。禁食和再投喂实验表明,与喂食组相比,金鱼肝脏中IGFBP-2的mRNA表达显著上调,再投喂后迅速恢复到正常水平。然而,金鱼下丘脑和垂体中IGFBP-2的mRNA表达对禁食不敏感。此外,下丘脑、垂体和肝脏中IGFBP-2的mRNA表达在进食前后变化,进食后2小时和4小时显著下调。这些结果表明,IGFBP-2 mRNA表达可能与金鱼的合成代谢和分解代谢相关,并受代谢因子调控。

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