《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (4): 858-865.doi: 10.16431/j.cnki.1671-7236.2018.04.003

• 生物技术 • 上一篇    下一篇

三穗麻鸭Cofilin2基因序列分析及其组织表达研究

华敏1, 贺欣薇1, 万润1, 程振涛1,2,3, 周碧君1,2,3, 文明1,2,3   

  1. 1. 贵州大学动物科学学院, 贵阳 550025;
    2. 贵州大学动物疫病研究所, 贵阳 550025;
    3. 贵州省动物疫病与兽医公共卫生重点实验室, 贵阳 550025
  • 收稿日期:2017-10-26 出版日期:2018-04-20 发布日期:2018-04-25
  • 通讯作者: 文明(1969-),博士,教授,研究方向:预防兽医学,E-mail:as.mwen@gzu.edu.cn E-mail:as.mwen@gzu.edu.cn
  • 作者简介:华敏(1990-),女,贵州普定人,硕士生,研究方向:预防兽医学,E-mail:1064063339@qq.com
  • 基金资助:

    国家自然科学基金项目(31260607、31560703);贵州省优秀青年科学人才培养计划项目(黔科合人字[2013]25号);贵州省百层次创新型人才项目(黔科合人才[2016]4009号);贵州省科技创新人才团队建设项目(黔科合人才团队[2015]4016号);贵州大学研究生创新基金项目(研农2017001)

Sequence Analysis of Cofilin2 Gene and Its Expression in Different Tissues of Sansui Ducks

HUA Min1, HE Xinwei1, WAN Run1, CHENG Zhentao1,2,3, ZHOU Bijun1,2,3, WEN Ming1,2,3   

  1. 1. College of Animal Science, Guizhou University, Guiyang 550025, China;
    2. Institute for Animal Diseases, Guizhou University, Guiyang 550025, China;
    3. Key Laboratory of Animal Diseases and Veterinary Public Health in Guizhou Province, Guiyang 550025, China
  • Received:2017-10-26 Online:2018-04-20 Published:2018-04-25

摘要:

为探究三穗麻鸭丝切蛋白2(Cofilin2)基因特征及其在鸭组织中的表达规律,试验根据GenBank中鸡Cofilin2基因序列设计特异性引物,利用RT-PCR方法扩增获得目的基因,利用生物信息学软件分析三穗麻鸭Cofilin2基因特征,实时荧光定量PCR方法检测Cofilin2基因在三穗麻鸭不同组织中的表达情况。结果显示,三穗麻鸭Cofilin2基因编码区大小为498 bp,可编码166个氨基酸;三穗麻鸭Cofilin2基因序列与鸡、猕猴、牛、小鼠、安大略鲑、人和猪相应序列的同源性分别为94.4%、91.6%、91.6%、87.6%、77.7%、70.5%和69.5%;系统进化树显示,Cofilin2基因在不同物种进化过程中高度保守;Cofilin2基因编码蛋白的二级结构由α-螺旋、延伸链、β-转角和无规则卷曲组成,三级结构呈弯曲螺旋结构;实时荧光定量PCR检测显示,三穗麻鸭Cofilin2基因在心脏、肝脏、脾脏、肺脏、肾脏、脑、胸腺、十二指肠、腿肌和法氏囊等组织器官中均有不同程度表达,其中心脏中表达量最高,法氏囊中表达量最低。本试验结果为三穗麻鸭抗病分子机理研究奠定了基础。

关键词: 三穗鸭; Cofilin2基因; 克隆; 实时荧光定量PCR

Abstract:

In order to analyze the characteristics of Cofilin2 gene and its expression in tissues of Sansui ducks,the specific primers were designed according to the sequence of Gullus Cofilin2 gene in GenBank,and amplified for Cofilin2 gene in Sansui ducks,then the Cofilin2 gene characteristic was analyzed using bioinformatic softwares,and the mRNA level of Cofilin2 gene in different tissues of Sansui ducks were detected with Real-time quantitative PCR.The results showed that the length of Cofilin2 gene was 498 bp in Sansui ducks,which encoded 166 amino acids.The identities of Cofilin2 gene in Sansui ducks were 94.4%,91.6%,91.6%,87.6%,77.7%,70.5% and 69.5% shared with that of Gallus,Macaca mulatta,Bos taurus,Mus musculus, Salmo salar,Homo sapiens and Sus scrofa,respectively.The phylogenetic tree analysis result showed that Cofilin2 gene maintained a highly conservative among different species.The secondary sructure of this encoding protein was consistes of alpha helix,extended strand,beta turn and random corn,and the curved spiral in tertiary structure.Real-time quantitative PCR detection results showed that there were different degrees expression of Cofilin2 gene in heart,liver,spleen,lung,kidney,brain,thymus,duodenum,leg muscle and bursa of Fabricius,the expression of Cofilin2 gene was the highest in heart,and that was the lowest in bursa of Fabricius.The results provided some basic data for the molecular mechanism of Sansui duck diseases.

Key words: Sansui ducks; Cofilin2 gene; cloning; Real-time quantitative PCR

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