遗传 ›› 2005, Vol. 27 ›› Issue (6): 908-914.

• 研究报告 • 上一篇    下一篇

昆明小鼠无毛基因cDNA全长序列的分子克隆

章金涛1~3;方盛国 ;2;王纯耀3;杜春燕3   

  1. 浙江大学生命科学学院濒危野生动物保护遗传学及繁殖生物学教育部重点实验室,杭州 310029
  • 收稿日期:2004-10-31 修回日期:2004-12-03 出版日期:2005-12-10 发布日期:2005-12-10
  • 通讯作者: 方盛国

Molecular Cloning of Full-long cDNA Sequences Encoding Hairless Gene in the Kunming Mouse

ZHANG Jin-Tao1~3, FANG Sheng-Guo1,2* ,WANG Chun-Yao3 , DU Chun-Yan3   

  1. College of Life Sciences, Zhejiang University,Hangzhou, 310029, China
  • Received:2004-10-31 Revised:2004-12-03 Online:2005-12-10 Published:2005-12-10
  • Contact: Shengguo Fang

摘要: 无毛基因是与皮肤和被毛结构有重要关联的核受体基因,编码一个锌指结构转录因子,是甲状腺激素受体的转录辅阻遏物,并参与毛发生长周期的调控,在维持毛囊及毛囊间上皮的增殖、分化、调亡的精致平衡中扮演重要角色。参考小鼠、人的无毛基因序列,用RT-PCR方法首次对昆明小鼠无毛基因的cDNA序列进行了克隆,获得了昆明小鼠无毛基因的全长4 014 bp cDNA序列(GenBank 登录号:AY547391),基因的CDS长度为3 546 bp。AY547391基因编码的蛋白质在GenBank中的序号为AAT45233,由1181个氨基酸组成。昆明小鼠与国外报导的小鼠、大鼠、猪、羊、猴、人等6种动物CDS 序列同源性分别为99.9%、94.4%、83.1%、78.1%、81.9%、82.1%,氨基酸同源性分别是99.9%、92.2%、81.7%、 70.8%、 79.9%、 80.1% 。这一结果反应了无毛基因在进化过程的高度保守性。通过Blast比较昆明小鼠无毛基因与GenBank 数据库中收集的Hr 基因的mRNA,发现了4个SNP和一个缺失突变,其中3个位点没有改变氨基酸残基的性质,两个位点有氨基酸改变并证实为多态性突变位点,研究结果为无毛基因的SNPs的数据库提供了新的信息。

关键词: 序列分析, 分子克隆, SNP, 小鼠, 无毛基因

Abstract: The mouse hairless gene is a crucial nuclear receptor gene associated with the structure of hair and skin. It encodes a putative zinc finger transcription factor, and is a transcriptional corepressor for thyroid hormone receptors. Hairless gene plays a critical role in maintaining the delicate balance between cell proliferation, differentiation, and apoptosis in the hair follicle as well as in the interfollicular epidermis, and is concerned with the control of hair growth cycling. This study was designed to clone and analyze the cDNA encoding Hr from Kunming mouse. The RT-PCR method was developed to clone the Hr cDNA. A full-length cDNA and CDS sequences of Kunming mouse were 4104bp and 3546bp, respectively, which has been accepted by GenBank (Accession Number: AY547391). Accession number of protein encoded by AY547391 sequence in GenBank is AAT45233 , composed of 1181 amino acid residues. The Hr cDNA of Kunming mouse was cloned and sequenced for the first time. The identities of CDS of Hr gene were 99.9%、94.4%、83.1%、78.1%、81.9% and 82.1% by homologous comparison among Kunming mouse and other six species, and that were 99.9%、92.2%、81.7%、 70.8%、 79.9% and 80.1% respectively in amino acid sequences. The results suggested a high degree of conservation and thus functional significance of the Hr gene among different mammalian taxons. The results derived from information searching by Blast program revealed that there were 4 SNP sites and one deletion mutation in the sequences of hairless gene mRNAs between Kunming mice and that collected in GenBank. Three SNP sites did not alter the related amino acids encoded. Two SNP proved to be polymorphism mutant sites with amino acids changes. The results provided new data for SNPs in Hr gene.

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