中国畜牧兽医

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山羊c-Myc基因的克隆及序列分析

安辰瑞,安铁洙,张秋婷,李昊,朴善花,王春生   

  1. (东北林业大学生命科学学院,黑龙江哈尔滨 150040)
  • 收稿日期:2013-10-25 出版日期:2014-04-20 发布日期:2014-05-27
  • 通讯作者: 王春生(1979―),男,黑龙江人,博士,副教授,硕士生导师,研究方向:体细胞重编程的分子机制。E-mail:wangchunsheng79@163.com
  • 作者简介:安辰瑞(1988―),男,辽宁人,硕士生,研究方向:体细胞重编程的分子机制。
  • 基金资助:

    中央高校基本科研业务费专项资金(DL13EA06-02);国家自然科学基金(31000990)。

Cloning and Sequence Analysis of c-Myc Gene in Goat

AN Chen-rui,AN Tie-zhu,ZHANG Qiu-ting,LI Hao,PIAO Shan-hua,WANG Chun-sheng   

  1. College of Life Science,Northeast Forestry University,Harbin 150040,China)
  • Received:2013-10-25 Online:2014-04-20 Published:2014-05-27

摘要: 为了克隆出山羊c-Myc基因的CDS区序列,本研究利用RT-PCR技术,参照GenBank报道的绵羊、牛、猪、马等的c-Myc基因CDS区序列设计出特异性引物,从山羊胚胎皮肤组织中扩增出山羊c-Myc CDS区序列并进行序列分析。结果表明,山羊c-Myc基因CDS区全长1320 bp (包括终止密码子),与绵羊c-Myc基因核苷酸序列(GenBank登录号:Z68501.1)比对仅有10处存在差异,其核苷酸序列同源性为99.17%,推导的氨基酸序列同源性为99.09%;与其他物种进行同源性分析结果显示,山羊c-Myc基因CDS区与猪、狼、人、大鼠、小鼠的核苷酸序列同源性分别为91.2%、92.0%、90.2%、86.9%、86.0%,推导的氨基酸序列同源性分别为93.6%、96.1%、92.3%、91.1%、89.6%;生物信息学软件分析结果表明,山羊c-Myc蛋白产物定位于细胞核并含有HLH结构域。本研究为进一步了解c-Myc基因的功能及探讨其在山羊体细胞重编程过程中的作用奠定了基础。

关键词: c-Myc基因; 基因克隆; 序列分析

Abstract: In order to clone sequence of the coding region of the goat c-Myc gene, this study designed specific primers by refering to c-Myc genes coding sequence reported in GenBank including sheep,cattle,pigs,horses,etc.,and then amplified c-Myc coding sequence from embryonic skin tissue of goats by using RT-PCR technique and had sequence analysis. The result indicated that CDS of goat c-Myc gene had 1320 bp length including the stop codon. For only 10 differences comparing to sheep c-Myc gene nucleotide sequence (GenBank accession number:Z68501.1),homology reached 99.17% and homology of deduced amino acid sequence reached 99.09%. Analysis of homology with other species showed that homology of goat c-Myc gene coding region nucleotide sequence was 91.2%,92.0%,90.2%,86.9%,86.0% with pigs,wolves,humans,rats,mice,and homology of deduced amino acid sequence was 93.6%,96.1%,92.3%,91.1%,89.6%. Using bioinformatics software analysis showed that goat c-Myc protein was localized in the nucleus and contained a HLH domain. This study further researched the function of c-Myc gene and laid the foundation of investigating the effect in the process of goat somatic cell reprogramming.

Key words: c-Myc gene; gene cloning; sequence analysis