北极狐GHR基因cDNA的克隆及序列分析-兽类学报.PDF

兽类学报,2010,30 (1):65 - 73 Acta Theriologica Sinica 北极狐 GHR 基因 cDNA 的克隆及序列分析 杜智恒 宁方勇 杨春山 白秀娟 (东北农业大学动物科技学院, 哈尔滨 150030) 摘要:本文根据狗 (AF133835 ) 的GHR 基因 cDNA 编码全序列设计了三对引物,利用 RT - PCR 方法克隆出北 极狐 GHR 基因编码区全长 cDNA 序列 (GenBank accession No EU304325 )。结果表明,北极狐 GHR 的 ORF 为 1 917 bp,编码638 个氨基酸的前体蛋白, 由18 个氨基酸的信号肽和620 个氨基酸的成熟肽组成。通过同源性比 较发现北极狐与狗的同源性最高,达到 98% 。另外,利用邻接法 (NJ 法) 构建的分子系统进化树聚类结果表 明,北极狐与狗先聚为一类,该聚类结果与传统的物种进化关系基本一致。另外,通过氨基酸对位序列比较发 现,北极狐 GHR 在氨基酸序列上存在明显的特异性,如45 和451 位分别为 A 和E,而其它物种均分别为T (大 鼠为K) 和A (牛羊为V, 鼠为T)。 关键词:北极狐;GHR 基因;克隆;序列分析 中图分类号:Q785 文献标识码:A 文章编号:1000 - 1050 (2010 )01 - 0065 - 09 cDNA cloning and sequence analysis of growth hormone receptor (GHR)from Arctic fox  DU Zhiheng,NING Fangyong,YANG Chunshan,BAI Xiujuan (College of Animal Science and Technology ,Northeast Agricultural University,Harbin 150030, China) Abstract :In the current study,the gene encoding growth hormone receptor (GHR)was cloned by RT - PCR from Arctic fox (Alopex lagopus)according to dog GHR gene (AF133835 ). Sequence analysis revealed that the open reading frame (ORF)of Arctic fox GHR is 1 917 bp,which encodes a638 - amino acid precursor containing a signal peptide of18 ami- no acids and a mature peptide of 620 Amino acids. Arctic fox GHR gene has highest similarity (98% )with dog GHR. The molecular phylogenetic tree was constructed by the NJ method based on GHR gene sequences. The result showed the Arctic fox and the dog were clustered into one branch,this result is consistent with traditional classification. In addition, Arctic fox GHR has its own characterization in its amino acid sequence compared to other mammal GHR sequences. For ex- ample,its amino acid residues at