高原世居藏族α、β珠蛋白编码基因的克隆与测序  

CLONING AND SEQUENCING OF α、β GLOBIN CODING GENES IN TIBETANS LIVING AT HIGH ALTITUDE

作　　者：董宏彬[1] 洪欣[1] 聂鸿靖[1] 肖忠海[1] 何贵祥[2] 黑沙汉 尹昭云[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所,天津300050 [2]西藏军区总医院医务处,拉萨850000 [3]青海省格尔木市人民医院,格尔木816000

出　　处：《中国应用生理学杂志》2005年第2期196-199,共4页Chinese Journal of Applied Physiology

基　　金：国家自然科学基因面上项目(30170354);重大项目(30393134)资助。

摘　　要：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。Aim: To explore the molecular biological mechanism of hemoglobin with high oxygen affinity in Tibetans by determining the sequence of globin cDNA in Tibetans living at high altitude. Methods: Total RNA was isolated from human bone marrow samples of three Tibetans who live in Qinghai-Tibet plateau. cDNA fragments coding for alpha and beta genes of human hemoglobin were obtained through RT-PCR and were ligated to plasmid pGEM-T easy vectors, and then the ligation liquid were transformed to Escherichia coli and cloned and sequenced. Nucleotide sequences were compared with GenBank data by BLAST method. Results: sequence of α globin cDNA in Tibetans were the same with the registering globin genes in the GenBank, and Hb Abruzzo(beta143(H21), His->Arg) gene mutation,a high oxygen affinity β globin mutation,was found in one Tibetan' beta goblin coding gene (CAC->CGC). Conclusion: This hemoglobin gene mutation may be associated with high altititude adaptation of Tibetans living at high altitude.

关 键 词：高原 藏族 珠蛋白 基因 HB Abruzzo 

分 类 号：R338[医药卫生—人体生理学] Q46[医药卫生—基础医学]