三穗麻鸭Cofilin2基因序列分析及其组织表达研究  

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

作　　者：华敏[1] 贺欣薇 万润[1] 程振涛 周碧君[1,2,3] 文明[1,2,3] HUA Min;HE Xinwei;WAN Run;CHENG Zhentao;ZHOU Bijun;WEN Ming(College of Animal Science,Guizhou University,Guiyang 550025,China;Institute for Animal Diseases,Guizhou University,Guiyang 550025,China;Key Laboratory of Animal Diseases and Veterinary Public Health in Guizhou Province,Guiyang 550025,China)

机构地区：[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

出　　处：《中国畜牧兽医》2018年第4期858-865,共8页China Animal Husbandry & Veterinary Medicine

基　　金：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)

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498bp,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。In order to analyze the characteristics of Cofilin 2 gene and its expression in tissues of Sansui ducks,the specific primers were designed according to the sequence of Gullus Cofilin 2 gene in GenBank,and amplified for Cofilin 2 gene in Sansui ducks,then the Cofilin 2 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 Cofilin 2 gene was 498 bp in Sansui ducks,which encoded 166 amino acids.The identities of Cofilin 2 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 Cofilin 2 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 Cofilin 2 gene in heart,liver,spleen,lung,kidney,brain,thymus,duodenum,leg muscle and bursa of Fabricius,the expression of Cofilin 2 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.

关 键 词：三穗鸭 Cofilin2基因 克隆 实时荧光定量PCR 

分 类 号：Q786[生物学—分子生物学]