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KMID : 0380220070400030426
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Journal of Biochemistry and Molecular Biology
2007 Volume.40 No. 3 p.426 ~ p.431
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Molecular Cloning of Hemoglobin Alpha-chain Gene from Pantholops hodgsonii, a Hypoxic Tolerance Species
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Yingzhong Yang
Droma Yunden
Guoen Jin
Zhenzhong Bai
Lan Ma
Haixia Yun
Yue Cao
Kubo Keishi
Rili Ge
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Abstract
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To investigate the possible mechanisms of high-altitude native animals in adapting to high altitude, we cloned hemoglobin alpha-chain (alpha-chain Hb) gene from Pantholops hodgsonii, an animal species that indigenously lives at elevations of 3700-5500 m on the Qinghai-Tibetan plateau. Using reverse transcription polymerase chain reaction (RT-PCR) technique, the alpha-chain Hb gene was amplified from total RNA in the liver of the Pantholops hodgsonii. TA cloning technique was used and the PCR product was cloned into pGEM-T vector. The DNA sequence of the gene was highly homologous with sheep (99.1%), goat (98.6%), cattle (95.6%) and human (86.5%). The alpha-chain Hb gene encoded a 142-amino acid protein that could be identified with the homology of alpha-chain Hb protein in sheep (98%), goat (96%), cattle (91%) and human (87%). However, 18 alternations were detected when compared with the alpha-chain Hb gene in human, and 2 in sheep. Moreover, the alterations of a117 GluAsp and &agr;132 AsnSer in important regions were noted in human and sheep, respectively. Phylogenetic analysis suggested that the structure of alpha-chain Hb was highly similar to that in sheep. This study provided essential information for elucidating the possible roles of hemoglobin in adapting to extremely high altitude in Pantholops hodgsonii.
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KEYWORD
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Adaptation, Alpha-chain Hb, Molecular cloning, Pantholops hodgsonii, Qinghai-Tibetan plateau
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