中文摘要：

克隆高原鼠兔脑红蛋白（Neuroglobin,NGB）基因编码区并检测其在成年高原鼠兔脑组织和其他组织中的表达,同时探讨高原鼠兔低氧适应的分子生物学机制。从高原鼠兔脑组织中提取总RNA,通过RT-PCR获得高原鼠兔NGB cDNA,将其与pGEM-TEasy载体连接,构建重组质粒,蓝白斑筛选阳性克隆并进行鉴定和测序 制备地高辛标记的RNA探针并采用原位杂交法（In suit hybridization,ISH）分析脑红蛋白基因在高原鼠兔脑组织中的分布 采用RT-PCR和蛋白印记（Westernblot）检测高原鼠兔不同组织中脑红蛋白的表达含量。将含有目的片段的阳性克隆经测序和Blast分析,显示其部分编码序列与GenBank中绵羊、大鼠等同源性很高（大于84%）,表明本实验所克隆的序列为脑红蛋白基因 原位杂交结果显示NGB在青藏高原土著动物高原鼠兔脑部分布较为广泛 高原鼠兔不同组织中都有NGB mRNA表达,NGB基因并不是中枢神经系统所特有的,睾丸和肾上腺也有较高的表达。NGB基因在高原鼠兔脑组织和其他组织中分布较为广泛,推测NGB mRNA可能在高原鼠兔机体较为广泛的区域中发挥着作用,同时为高原低氧适应相关基因的研究提供了实验依据。

英文摘要：

For identification of the neuroglobin genes coding sequences molecular cloning and examination of the tissues expression spectrums and showing the hypoxic adaptations mechanisms in Plateau pika（Ochotona curzoniae）.Extracting the total RNA, cloning neuroglobin coding sequences cDNA with reverse-transcription RT-PCR, capturing and confirming the certain sequences with DNA sequencing.Examination the tissues mRNA expressions spectrums of neuroglobin with in suit hybridization ISH technology.In addition, semi-quantitative RT-PCR and western-blot technologies were being used to analysis of the relative neuroglobin mRNA and protein expression amounts in various tissues with Plateau pika.Cloning and sequencing results confirmed it was the neruoglobin gene coding sequence of Plateau pika with Blast analysis; and the ISH results exhibited the neuroglobin gene expressions spectrums indicate widely distributed with higher amount in brain tissues of Plateau pika.Additionally, neuroglogin mRNA also expressed in the other tissues such as testis and adrenal glands besides brain tissues. We can speculate that NGB might play an important role in the adaptations of the Plateau pika under the high altitude environment. Furthermore, native Tibetan species neuroglobin genes will also present fundamental evidence to investigate high altitude adaptable related genetics study in the future.