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藏羚羊低氧诱导因子1α基因的克隆与组织表达

刘芳, 乌仁塔娜, 马兰, 杨应忠, 格日力^*

青海大学医学院高原医学研究中心，西宁 810001

摘要

为探讨低氧诱导因子1α (hypoxia inducible factor 1 alpha, HIF-1α)在藏羚羊(Pantholops hodgsonii)高原低氧适应机制中的作用，采用RT-PCR和RACE技术克隆藏羚羊HIF-1α基因的cDNA序列，同时采用real-time PCR和Western blot方法检测藏羚羊、藏系绵羊、平原绵羊HIF-1α的组织特异性表达，并进行比较分析。序列分析结果表明，克隆所得的cDNA序列为3 664 bp，包含2 471 bp的开放阅读框和1 191 bp的3’UTR区。它的编码区序列和预测的氨基酸序列与其它哺乳动物HIF-1α的相似性超过87%，其中与牛相似性最高，达到98%以上，表明该序列为藏羚羊HIF-1α基因的cDNA序列。基因表达检测结果显示，HIF-1α在藏羚羊的肺、心肌、骨骼肌中均有表达，其中肺的表达量最高，表现出明显的组织差异性。进一步研究表明，藏羚羊和藏系绵羊的HIF-1α蛋白在上述三种组织中的表达量均高于平原绵羊，并且HIF-1α mRNA和蛋白在藏羚羊心、肺和骨骼肌中的表达量高于藏系绵羊。提示HIF-1α的低氧特异表达是藏羚羊高原低氧适应的分子基础。

关键词： 低氧诱导因子1α; 藏羚羊; 基因克隆; 基因表达; 低氧适应性

分类号：Q78

[Genetic cloning and expression of hypoxia inducible factor 1 alpha in high altitude hypoxic adaptation species Tibetan antelope (Pantholops hodgsonii).] [Article in Chinese]

LIU Fang, Wuren Tana, MA Lan, YANG Ying-Zhong, GE Ri-Li^*

Research Center of High Altitude Medicine, Qinghai University Medical College, Xining 810001, China

Abstract

In order to investigate the role of the hypoxia inducible factor 1 alpha (HIF-1α) in the adaptation mechanism to high altitude hypoxia, the cloning of the HIF-1α gene cDNA of Tibetan antelope (Pantholops hodgsonii), using RT-PCR and RACE, was applied, and the comparative analysis of the tissue-specific expressions of HIF-1α among Tibetan antelope, Tibetan sheep and plain sheep was performed using real-time PCR and Western blot. The sequence analysis indicated that the cDNA sequences acquired by cloning from the HIF-1α gene of Tibetan antelope comprised a 2 471-bp open reading frame (ORF) and a 1 911-bp 3’UTR. The similarity between its coding sequence, predicted amino acid sequence and HIF-1α of other mammals exceeded 87%, in which the similarity with cow was up to more than 98%, which showed that this sequence was the cDNA of HIF-1α of Tibetan antelope. The results of real-time PCR and Western blot showed that expressions of HIF-1α mRNA and protein appeared in Tibetan antelope’s lung, cardiac muscle and skeletal muscle, with the highest expression in lung. HIF-1α mRNA and protein had obvious differential expression in these tissues. Further research showed that Tibetan antelope and Tibetan sheep possessed higher expressions of HIF-1α protein in the three tissues above-mentioned compared with plain sheep, and the expressions of HIF-1α mRNA and protein in Tibetan antelope’s lung, cardiac muscle and skeletal muscle were higher than those of Tibetan sheep. It illustrates that the hypoxic HIF-1α-specific expression is one of the molecular bases of high altitude hypoxia adaptation in Tibetan antelope.

Key words: hypoxia inducible factor 1 alpha; Tibetan antelope; genetic cloning; genetic expression; hypoxia adaptation

收稿日期：2011-05-19　　录用日期：2011-08-04

通讯作者：格日力　　E-mail: geriligao@hotmail.com

引用本文：

刘芳, 乌仁塔娜, 马兰, 杨应忠, 格日力. 藏羚羊低氧诱导因子1α基因的克隆与组织表达[J]. 生理学报 2011; 63 (6): 565-573.

LIU Fang, Wuren Tana, MA Lan, YANG Ying-Zhong, GE Ri-Li. [Genetic cloning and expression of hypoxia inducible factor 1 alpha in high altitude hypoxic adaptation species Tibetan antelope (Pantholops hodgsonii).] [Article in Chinese]. Acta Physiol Sin 2011; 63 (6): 565-573 (in Chinese with English abstract).