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高原鼠兔ob 基因的组织表达特征

杨洁, 赵新全,郭松长, 李红阁, 祁得林, 王德鹏, 曹俊虎   

  1. 中国科学院西北高原生物研究所,西宁
  • 出版日期:2007-03-17 发布日期:2008-07-07

The tissue expression character of ob mRNA in plateau pika (Ochotona curzoniae)

YANG Jie, ZHAO Xinquan, GUO Songchang, LI Hongge, QI Delin,WANG Depeng,CAO Junhu   

  • Online:2007-03-17 Published:2008-07-07

摘要: ob 基因编码的leptin 蛋白在调节生物体能量平衡中起到重要作用。本研究应用Taqman 探针real time PCR 技术对高原鼠兔ob 基因的组织分布进行检测。通过提取不同组织总RNA,经DNase I 消化后,用随机引物进行反转录合成cDNA,采用特异性Taqman 探针和引物分别对ob 基因及β-actin 基因进行实时定量PCR 扩增,对不同组织中ob 基因和β-actin 基因的初始拷贝数之比进行比较。结果表明ob 基因在脑、心脏、肺、肝脏、脾脏、肾脏、骨骼肌、脂肪组织中均有表达,其中以白色脂肪组织中ob 基因表达量最高,其次为心脏和肺,表达量最低的是肝脏和肾脏。

关键词: 高原鼠兔, ob 基因, Taqman 探针, Real time PCR, 组织分布

Abstract: Leptin, the production of the ob gene, plays an important role in the regulation of energy homeostasis. In this study, we detected the tissue expression character of ob gene in plateau pika (Ochotona curzoniae) using the method of real time PCR (Taqman probe). Total RNA was extracted from eight issues includingbrain, heart, lung, liver, spleen, kidney, muscle and white adipose of plateau pika. After treated with DNase I, the total RNA was reverse transcribed with random primer to synthesize cDNA. Then the ob gene and β-actin gene were amplified with pika-specific Taqman probes and primers by real time PCR. The pika leptin mRNA in different tissues were normalized with β-actin mRNA to compensate for variations in input RNA amounts.Our results showed that ob mRNA was expressed in all of eight tissues, and tissular specificity was displayed with the highest in white adipose, next in heart and lung, and much less in kidney and liver.

Key words: Ob gene, Plateau pika (Ochotona curzoniae), Real time PCR, Taqman probe, Tissue distribution