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Genetic cloning and expression of neuroglobin in high altitude hypoxic ad- aptation species——Plateau pika (Ochotona curzoniae) |
BAI Zhenzhong,HAN Shufen,YANG Yingzhong,CAO Yue,MA Lan,LIU Shou,GE Rili |
Research Center for High Altitude Medicine,Qinghai University Medical School,Xining 810001,China |
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Abstract 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.
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