《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (5): 1290-1298.doi: 10.16431/j.cnki.1671-7236.2019.05.005

• 生物技术 • 上一篇    下一篇

哈萨克羊INHβA基因克隆及生物信息学分析

解津刚1, 巴合提·博代1, 热衣扎别克·哈德坎2, 吾热力哈孜·哈孜汗1   

  1. 1. 石河子大学动物科技学院, 石河子 832000;
    2. 哈萨克国立农业大学动物生物资源系, 阿拉木图 050010
  • 收稿日期:2018-07-30 出版日期:2019-05-20 发布日期:2019-05-20
  • 通讯作者: 吾热力哈孜·哈孜汗 E-mail:1508217366@qq.com
  • 作者简介:解津刚(1997-),男,新疆乌鲁木齐人,硕士生,研究方向:动物遗传育种与繁殖,E-mail:838245737@qq.com
  • 基金资助:

    兵团国际科技合作项目(2016AH001)

Cloning and Bioinformatics Analysis of INHβA Gene in Kazakh Sheep

XIE Jingang1, Baheti Bodai1, Kadyken Rizabek2, Wurelihazi Hazihan1   

  1. 1. College of Animal Science, Shihezi University, Shihezi 832000, China;
    2. Department of Technology and Biological Resources, Kazakh National Agrarian University, Almaty 050010, Kazakhstan
  • Received:2018-07-30 Online:2019-05-20 Published:2019-05-20

摘要:

试验旨在克隆哈萨克羊抑制素βA (inhibin beta A,INHβA)基因序列,并探究其序列特征及编码蛋白的结构和功能。根据GenBank中绵羊INHβA基因序列(登录号:NM_001009458.1)设计1对特异性引物,通过RT-PCR方法对哈萨克羊INHβA基因进行扩增,将获得的INHβA基因片段插入到pMD19-T载体中进行克隆测序,并结合生物信息学方法预测和分析其核苷酸序列、氨基酸序列、蛋白跨膜、蛋白修饰位点及二级结构、三级结构模型等。结果显示,哈萨克羊INHβA基因长1 278 bp,编码425个氨基酸。哈萨克羊INHβA基因与绵羊、牛、野猪、小鼠、人、大鼠、猫、兔、马的同源性分别为98.6%、97.7%、90.4%、87.9%、91.1%、88.2%、91.8%、89.8%和91.8%,表明INHβA基因在不同物种之间具有较高的保守性。INHβA蛋白分子式为C2072H3325N603O628S26,分子质量为47.57 ku,理论等电点(pI)为7.87,不稳定系数为66.16,脂溶指数为78.47,亲水值为-0.507。INHβA是一种碱性不稳定的亲水性脂溶性蛋白,没有跨膜结构,含有信号肽。二级结构预测显示,INHβA蛋白α-螺旋、β-转角、无规则卷曲、延伸链分别占22.59%、4.47%、54.12%和18.82%。三级结构预测显示,INHβA蛋白以α-螺旋为主,是含有8种蛋白修饰位点的β-桶状蛋白。本试验结果为深入研究哈萨克羊INHβA蛋白功能及探讨INHβA基因对提高哈萨克羊繁殖力的影响提供参考数据。

关键词: 哈萨克羊; INHβ A基因; 克隆; 序列分析

Abstract:

This study was aimed to clone the sequence of inhibin beta A (INHβA) in Kazakh sheep,and explore the sequence characteristics of INHβA gene and the structure and function of its encoded protein.One pair of specific primers was disigned based on the sequence of sheep INHβA gene (accession No.:NM_001009458.1) in GenBank,the sequence of INHβA gene in Kazakh sheep was amplified by RT-PCR.INHβA gene was inserted into pMD19-T vector for cloning and sequencing,its nucleotide sequence,amino acids sequence,protein transmembrane,protein modification site and secondary structure,tertiary structure model were predicted and analyzed by bioinformatics methods.The results showed that INHβA gene in Kazakh sheep was 1 278 bp in length and encoded 425 amino acids.The homology of INHβA gene in Kazakh sheep with Ovis aries,Bos taurus,Sus scrofa,Mus musculus,Homo sapiens,Rattus norvegicus,Felis catus,Rytolagus cuniculus and Equus caballus was 98.6%,97.7%,90.4%,87.9%,91.1%,88.2%,91.8%,89.8% and 91.8%,respectively,indicating that INHβA gene was highly conserved among different species.The molecular formula of INHβA protein was C2072H3325N603O628S26,the molecular weight was 47.57 ku,the theoretical isoelectric point (pI) was 7.87,the instability coefficient was 66.16,the fat solubility index was 78.47,and the hydrophilic value was -0.507.The INHβA protein was an alkaline labile hydrophilic liposoluble protein containing a signal peptide,with no transmembrane structure.The secondary structure prediction showed that alpha helix,beta turn,random coil,and extended chain of INHβA protein accounted for 22.59%,4.47%,54.12% and 18.82%,respectively.The tertiary structure prediction showed that the spatial structure of the protein was mainly alpha helix,and contained β-barrel structures of 8 protein modification sites.This results provided a data reference for the later study of the function of INHβA protein and the effect of INHβA gene on improving the fecundity in Kazakh sheep.

Key words: Kazakh sheep, INHβA gene, cloning, sequence analysis

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