藏羚羊α-珠蛋白基因编码区的克隆及同源性分析  

Cloning and homologous analysis of coding region of α-globin gene from Tibetan antelope

作　　者：杨应忠[1] 白振中[1] 靳国恩[1] 马兰[1] 韵海霞[1] 格日力[1] 

机构地区：[1]青海大学医学院高原医学研究中心,青海西宁810001

出　　处：《细胞与分子免疫学杂志》2007年第7期620-622,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30393133);青海省重点科技攻关项目(2003-N-120)

摘　　要：目的:克隆藏羚羊α-珠蛋白基因编码区,并进行同源性分析。方法:从藏羚羊肝组织中提取总RNA,通过RT-PCR获得藏羚羊α-珠蛋白cDNA,将其与pGEM(-T Easy载体连接,构成重组质粒,并转化大肠杆菌TOP10扩增培养后,鉴定阳性质粒并进行测序。将测序的结果与NCBI数据库进行同源性比较。结果:获得α-珠蛋白编码区cDNA序列,提交GenBank数据库并取得注册号为DQ650713。与绵羊α-珠蛋白基因相比较,其在16、53、132、134处出现核苷酸密码子突变(GAC→GGC)、(TCG→TCC)、(AGC→GGC)、(AAC→AGC)。推导的氨基酸序列比较发现,132位的天冬酰胺酸(Asn)突变为丝氨酸(Ser),134位的丝氨酸(Ser)突变为甘氨酸(Gly);与人类的α-珠蛋白相比较,有19处的氨基酸发生改变。结论:成功地克隆出藏羚羊α-珠蛋白基因编码区,为高原低氧适应相关基因的研究提供了实验依据。AIM： To clone and analyze the encoding region of α-globin gene from Tibetan antelope. METHODS： Total RNA was isolated from an adolescent Tibetan antelope liver, and Tibetan antelope α-globin gene was amplified by RT-PCR. The PCR product was cloned into pGEM-T vector and sequenced. Nucleotide sequences were compared with GenBank data by Blast method. RESULTS. The encoding region of α-globin gene of Tibetan antelope was obtained and deposited in GenBank as accession number DQ650713. Compared with sheep α-chain, alterations in important regions could be noted： a132 Asn→Ser, a134 Ser→Gly; but 19 differences were detected when compared with that of human. Phylogenetic analysis showed that the encoding region of α-globin gene of Tibetan antelope was most likely close to that of sheep and goat. CONCLUSION： The encoding region of gene Tibetan antelope α-globin gene is successfully cloned, which provides basic information for elucidating the possible role of hemoglobin in high altitude adaptation of Tibetan antelope.

关 键 词：高原适应 藏羚羊 Α-珠蛋白 基因克隆 同源性分析 

分 类 号：Q786[生物学—分子生物学]