藏羚羊低氧诱导因子1α基因的克隆与组织表达  

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

作　　者：刘芳[1] 乌仁塔娜[1] 马兰[1] 杨应忠[1] 格日力[1] 

机构地区：[1]青海大学医学院高原医学研究中心,西宁810001

出　　处：《生理学报》2011年第6期565-573,共9页Acta Physiologica Sinica

基　　金：supported by the National Basic Research Development Program of China(No.2012CB518200);the National Natural Science Foundation of China(No.31161861)

摘　　要：为探讨低氧诱导因子1α(hypoxia inducible factor1alpha,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3664bp,包含2471bp的开放阅读框和1191bp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-1αmRNA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。In order to investigate the role of the hypoxia inducible factor 1 alpha （HIF-1a） m the aaaptatlon mecnanlsm to nxgn amtude hypoxia, the cloning of the HIF-1 a 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-1a 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-lcL 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-1a 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- 1a of Tibetan antelope. The results of real-time PCR and Western blot showed that expressions of HIF-1a mRNA and protein appeared in Tibetan antelope＇s lung, cardiac muscle and skeletal muscle, with the highest expression in lung. HIF-1a mRNA and protein had obvious differential expression in these tissues. Further research showed that Tibetan antelope and Tibetan sheep possessed higher expressions of HIF-1a protein in the three tissues above-mentioned compared with plain sheep, and the expressions of HIF-1a 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-1a-specific expression is one of the molecular bases of high altitude hypoxia adaptation in Tibetan antelope.

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[生物学—分子生物学]