《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (3): 619-627.doi: 10.16431/j.cnki.1671-7236.2017.03.001

• 生物技术 •    下一篇

树鼩Tssk6基因cDNA克隆、组织表达谱及生物信息学分析

李元吉1, 杨明华1, 刘芳1, 朱方方1, 和绍禹2, 匡海鸥2, 李亚辉2   

  1. 1. 云南农业大学动物科学技术学院, 昆明 650201;
    2. 云南农业大学食品科学技术学院, 昆明 650201
  • 收稿日期:2016-09-12 出版日期:2017-03-20 发布日期:2017-03-21
  • 通讯作者: 李亚辉 E-mail:kmliyh@163.com
  • 作者简介:李元吉(1989-),男,河南叶县人,硕士生,研究方向:动物生殖生物学,E-mail:yuanjilee@hotmail.com;杨明华(1967-),女,云南陇川人,硕士,实验师,研究方向:动物分子营养与代谢调控,E-mail:yangmh85@hotmail.com
  • 基金资助:

    国家自然科学基金(31460308);云南中科灵长类生物医学重点实验室开放基金(云中科字2016-008号);国家自然科学基金专项基金(31040055)

cDNA Cloning, Tissue Expression Profiles and Bioinformatical Analysis of Tssk6 Gene in Tree Shrews

LI Yuan-ji1, YANG Ming-hua1, LIU Fang1, ZHU Fang-fang1, HE Shao-yu2, KUANG Hai-ou2, LI Ya-hui2   

  1. 1. Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;
    2. College of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, China
  • Received:2016-09-12 Online:2017-03-20 Published:2017-03-21

摘要:

本研究旨在克隆出树鼩Tssk6基因cDNA的全长序列,阐明其组织表达谱及基本生物信息学特征。以GenBank中收录的树鼩Tssk6基因mRNA预测序列为参考设计特异性引物,对树鼩Tssk6基因的cDNA全长序列进行克隆,用实时荧光定量PCR检测该基因在树鼩不同组织中的表达情况,并对cDNA序列的CDS区核苷酸序列与蛋白质结构进行了生物信息学分析。结果显示,树鼩Tssk6基因cDNA的CDS区序列长度为822 bp,编码273个氨基酸;Tssk6基因仅表达于树鼩睾丸组织;对树鼩和其他8种哺乳动物Tssk6基因cDNA的CDS区核苷酸序列构建的系统进化树显示,树鼩与人、黑猩猩、恒河猴3种灵长类动物亲缘关系较近,与小鼠、大鼠、金仓鼠3种啮齿类动物亲缘关系较远。本研究为进一步研究树鼩Tssk6基因的功能奠定了基础。

关键词: 树鼩; Tssk6基因; cDNA; 克隆; 生物信息学; 实时荧光定量PCR

Abstract:

The purpose of this study was to clone the full-length cDNA of tree shrew's Tssk6 gene, to elucidate its tissue expression profiles and fundamental bioinformatical characteristics. In the present study, the full-length cDNA of tree shrew's Tssk6 gene was cloned with specific primers on the basis of predicted mRNA sequence of tree shrew's Tssk6 gene in GenBank, the mRNA expression of Tssk6 gene in various tree shrew's tissues was investigated via Real-time PCR. In addition, the bioinformatics on nucleotide sequence of CDS region of Tssk6 cDNA in tree shrew as well as putative protein structure had been analyzed. The results showed that:The CDS region of tree shrew's Tssk6 cDNA contained 822 bp encoding 273 amino acids; Tssk6 gene specifically expressed in tree shrew's testis; Based on the molecular phylogenetic tree constructed with CDS of Tssk6 cDNA among tree shrew and other eight mammalian species, it was inferred that tree shrew was genetically close to primates like human, chimpanzee and rhesus monkey, while it was genetically far to rodents like mouse, rat and golden hamster. This study would lay a foundation for the further study of Tssk6 gene function in tree shrew.

Key words: tree shrew; Tssk6 gene; cDNA; cloning; bioinformatics; Real-time PCR

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