牦牛TRIB3基因克隆、生物信息学及组织表达差异分析  

Cloning of TRIB3 gene in yak and analyzing its bioinformatics and tissue expression difference

作　　者：MEHRAN Sidra 董宝霞 荆海霞[1] MEHRAN Sidra;DONG Baoxia;JING Haixia(College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学农牧学院,青海西宁810016

出　　处：《畜牧与兽医》2024年第5期1-9,共9页Animal Husbandry & Veterinary Medicine

基　　金：青海省科技厅项目(2019-ZJ-7025)。

摘　　要：旨在克隆牦牛毛球族同源蛋白3(TRIB3)基因并进行生物信息学分析,检测其在牦牛各组织中的表达情况,为后续探究该基因在牦牛中的功能提供理论依据。试验采集牦牛乳腺组织并以其cDNA为模板,克隆牦牛TRIB3基因序列,通过生物信息学分析了解其生物学特征和结构,运用荧光定量PCR(RT-qPCR)技术检测TRIB3基因在牦牛心脏、肝脏、肺脏、骨骼肌、乳腺及脂肪组织中的表达情况。结果显示:牦牛TRIB3基因CDS区全长为1074 bp,共编码357个氨基酸,为不稳定性亲水蛋白;同源性对比结果显示,牦牛TRIB3编码氨基酸与野牦牛的同源性最高;系统进化树显示,野牦牛和野牛与牦牛TRIB3基因亲缘关系最近,最远是小鼠;TRIB3蛋白属于不稳定亲水性蛋白,主要存在于细胞核中,不含信号肽和跨膜结构域;TRIB3蛋白二级结构含有α-螺旋、延伸链、β-转角、无规则卷曲,分别占35.85%、9.52%、448%和50.14%;蛋白互作分析结果显示,TRIB3与组成型光形态建成蛋白1(COP1)、过氧化物酶体增殖物激活受体α(PPARα)、CCAAT增强子结合蛋白β(CEBPβ)、激活转录因子3(ATF3)、DNA损伤诱导转录因子3(DDIT3)、丝氨酸/苏氨酸蛋白激酶2(AKT2)、AKT3和Twist相关蛋白1(TWIST1)之间均有紧密联系。RT-qPCR检测发现TRIB3基因在牦牛的不同组织中均有表达,乳腺组织中的表达量最高。研究结果为进一步探究TRIB3基因在牦牛乳腺中的作用和其生物功能提供一定的理论数据。This study was to clone the yak tribbles homologous protein 3(TRIB3)gene,and to analyze its bioinformatics and detect its ex-pression difference in yak tissues,so as to lay a foundation for further research on the function of this gene in yak.In the experiment,samples of breast tissue were collected from yak and the cDNA of the tissue was used as a template to clone the yak TRIB3 gene sequence,and the bi-ological characteristics and structure of the sequence were analyzed through bioinformatics.Then,RT-qPCR was used to detect the expres-sion of TRIB3 genes in yak heart,liver,lungs,skeletal muscle,breast and adipose tissue.The results showed that the total length of the CDS region of the TRIB3 gene in yaks was 1074 bp,encoding a total of 357 amino acids,and it was an unstable hydrophilic protein.The re-sults of homology comparison showed that the amino acid encoding TRIB3 of yak was in the highest homology with that of wild yak.The phylo-genetic tree showed that wild yak and buffalo had the closest genetic relationship with the TRIB3 gene of yak,and the farthest was mouse.TRIB3 protein was an unstable hydrophilic protein,which mainly existed in the nucleus and did not contain signal peptide or transmembrane domain.The secondary structure of the TRIB3 protein containedαhelix,extended chain,βturn,and random coil,accounting for 35.85%,9.52%,4.48%,and 50.14%,respectively.The results of protein interaction analysis showed that TRIB3 was closely related to COP1,PPARA,CEBPβ,ATF3,DDIT3,AKT2,AKT3,and TWIST1 proteins.RT-qPCR detection revealed that the TRIB3 gene expressed in dif-ferent tissues of yak,and the highest expression level of the TRIB3 gene was in yak breast tissue.These results provided theoretical data for further exploring the role of the TRIB3 gene in yak mammary gland and its biological function.

关 键 词：牦牛 毛球族同源蛋白3 基因克隆 生物信息学分析 组织表达 

分 类 号：S852[农业科学—基础兽医学]