生物技术通报 ›› 2017, Vol. 33 ›› Issue (2): 97-101.doi: 10.13560/j.cnki.biotech.bull.1985.2017.02.014

• 研究报告 • 上一篇    下一篇

齐口裂腹鱼白介素-1β基因的克隆及表达谱

段荟芹1, 王利2   

  1. 1. 西南民族大学青藏高原研究院,成都 610041;
    2. 西南民族大学生命科学与技术学院,成都 610041
  • 收稿日期:2016-06-16 出版日期:2017-02-26 发布日期:2017-02-08
  • 作者简介:段荟芹,女,硕士生,研究方向:动物遗传学;E-mail:917077886@qq.com
  • 基金资助:
    四川省科技支撑项目(2016NZ0044),西南民族大学研究生学位点建设项目(2016XWD-SO71007)

Cloning and Expression Profile of Interleukin-1β Gene in Schizothorax prenanti

DUAN Hui-qin1, WANG Li2   

  1. 1. Institute of Qinghai-Tibet Plateau,Southwest University for Nationalities,Chengdu 610041;
    2. College of Life Science and Technology,Southwest University for Nationalities,Chengdu 610041
  • Received:2016-06-16 Published:2017-02-26 Online:2017-02-08

摘要: 探讨齐口裂腹鱼白介素-1β(Interleukin-1β,IL-1β)基因的基本特点。用RT-PCR方法以齐口裂腹鱼脾脏cDNA作为模板扩增IL-1β基因,用生物信息学软件分析其序列,用RT-PCR方法检测IL-1β基因在齐口裂腹鱼6种组织的表达情况。克隆获得的IL-1β序列长1 252 bp,共编码276个氨基酸。GenBank 序列号为KU886235。IL-1β蛋白相对分子量为31.25 kD,等电点5.45,预测为亲水性蛋白。二级结构主要结构元件为随机卷曲和延伸链,有4个N-糖基化位点和16个磷酸化位点。NJ法系统进化树显示,齐口裂腹鱼与鲤鱼、鲫鱼的亲缘关系最近。组织表达分析显示,IL-1β基因主要在脾脏和肾脏中表达。这为深入研究鱼IL-1β的生物学功能提供理论依据。

关键词: 齐口裂腹鱼, 白介素-1β, 克隆, 蛋白质分析, 基因表达

Abstract: This work aims to characterize interleukin-1β(IL-1β)gene of Schizothorax prenanti. First,RT-PCR was used to amplify IL-1β gene with cDNA of S. prenanti spleen as template. Then,the IL-1β gene sequence was analyzed with various bioinformatics tools,and the expressions in 6 tissues of S. prenanti were detected used RT-PCR. As results,the length of IL-1β gene was 1 252 bp,and it encoded 276 amino acids. Its GenBank accession number was KU886235,the molecular weight of IL-1β was 31.25 kD,and pI was 5.45. It was induced as a hydrophilic protein,and its secondary structure mainly consisted of random coil and extended strand. There were 4 N-glycosylation sites and 16 phosphorylation sites in this protein. Phylogenetic relationships revealed that S. prenanti presented a closest genetic relationship with Cyprinus carpio and Carassius auratus. The analysis of tissue expression indicated that IL-1β gene was mainly expressed in the spleen and kidney. It provides theoretical evidence for further studying the biological function of IL-1β gene.

Key words: Schizothorax prenanti, interleukin-1β, cloning, protein analysis, gene expression