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目的:通过对高原世居藏族α、β珠蛋白编码基因的分析,探讨藏族Hb高氧亲合力的分子机制。方法:高原现场采集健康成年男性藏族人骨髓样品,提取总RNA,通过逆转录聚合酶链反应(RT PCR)获得人α和β珠蛋白的cDNA,与PGEM-TEasy质粒连接后,将α和β珠蛋白的cDNA转化JM109大肠杆菌中扩增培养,经酶切鉴定后测序,结果与NCBI数据库进行同源性比较。结果:藏族人α珠蛋白的cDNA与NCBI数据库登录的人cDNA序列相同,没有突变位点。一例藏族人β珠蛋白143位密码子发生了氧亲和力增高的碱基突变(CAC>CGC),其对应的氨基酸由His变为Arg(即HbAbruzzo)。结论:藏族人高氧亲和力变种的发现,为今后高原低氧适应相关基因的研究提供了线索。
OBJECTIVE: To investigate the molecular mechanism of Tibetan Hib (Hb) hyperoxygenation by analyzing the gene encoding α, β globin gene in native Tibetan Plateau. METHODS: Bone marrow samples of healthy adult male Tibetans were collected on the plateau. Total RNA was extracted and cDNA of human α and β globin was obtained by reverse transcription polymerase chain reaction (RT PCR). After being ligated with PGEM-TEasy plasmid, α and β The cDNA of globin was transformed into E.coli JM109 for amplification. After restriction enzyme digestion and sequencing, the results were compared with NCBI database. Results: Tibetan human α-globin cDNA was identical to the human cDNA sequence registered in the NCBI database, with no mutation sites. One case of Tibetan beta globin 143 codon with a high oxygen base mutation (CAC> CGC), the corresponding amino acid change from His to Arg (HbAbruzzo). CONCLUSION: The discovery of Tibetan hyperoxygenin variants provides clues for the study of hypoxia adaptation-related genes in the future.