高原鼠兔血红素氧合酶1的基因克隆及序列分析

 摘 要] 目的:克隆高原鼠兔血红素氧合酶1（heme oxygenase-1，HO-1）基因cDNA的全长序列，并分析其序列特征，为进一步揭示高原鼠兔低氧适应的分子机制提供有益参考。方法:从高原鼠兔肝组织中提取总RNA，利用逆转录聚合酶链反应（RT-PCR）技术和cDNA末端快速克隆（RACE）技术扩增出HO-1基因cDNA全长序列并进行测序，测序结果采用生物信息学的方法进行分析。结果:克隆所得鼠兔HO-1基因cDNA全长片段大小为1 466 bp，与预期一致，编码区长度为873   bp，编码290个氨基酸（GenBank登录号为：JX035934）；序列分析结果显示，核苷酸序列和推测的氨基酸序列相似性比较，与兔、人、牛、小鼠、大鼠、猪和马的HO-1核苷酸序列的同源性分别为89%、87%、85%、79%、84%、85%和85%，与兔、人、牛、小鼠、大鼠、猪和马的HO-1氨基酸序列的同源性分别为89%、85%、84%、80%、79%、82%和67%,显示出高度保守性。构建的基于氨基酸序列的分子系统进化树聚类结果表明，高原鼠兔与兔的进化距离最近。结论: 本实验首次成功克隆出高原鼠兔HO-1基因cDNA全长序列，为从低氧细胞保护角度，进一步探讨青藏高原土著物种适应高原的分子生物学机制研究提供实验依据。

Cloning and sequencing of heme oxygenase-1 gene from Ochotona curzo-niae

ABSTRACT] AIM: To clone and analyze the heme oxygenase-1 (HO-1) gene from Ochotona curzoniae (plateau pika). METHODS: The cDNA of HO-1 was cloned by RT-PCR and rapid amplication of cDNA ends (RACE) from the liver of Ochotona curzoniae. The bioinformatic analysis of HO-1 gene was performed. RESULTS: The cDNA of HO-1 gene in Ochotona curzoniae was obtained. The data of the sequence was deposited into GenBank and the accession number is JX035934. The full length of cDNA was 1 466 bp, including 873 bp encoding sequence (290 amino acids). Homology comparison showed that the DNA sequence of the HO-1 gene was highly homologous with Oryctolagus cuniculus (89%), Homo sapiens (87%), Bos taurus (85%), Mus musculus (79%), Rattus norvegicus (84%), Sus scrofa (85%) and Equus caballus (85%), and the amino acid sequence of HO-1 was identified with the homology of 89%, 85%, 84%, 80%, 79%, 82% and 67%, respectively. Phylogenetic analysis suggested that the structure of HO-1 was highly similar to that in Oryctolagus cuniculus. CONCLUSION: The HO-1 gene of Ochotona curzoniae was successfully cloned and provides essential information for elucidating the possible roles of HO-1 in adapting of Ochotona curzoniae to extremely high altitude.