黔北麻羊GPx3基因克隆、生物信息学分析及在不同性腺组织中的表达分析  

Cloning,Bioinformatics Analysis and Expression Analysis of GPx3 Gene in Different Gonad Tissues of Qianbei Ma Goat

作　　者：付开斌 陈祥[1,2] 周志楠 张艳[1,2] 惠茂茂[1,2] 洪磊 FU Kaibin;CHEN Xiang;ZHOU Zhinan;ZHANG Yan;HUI Maomao;HONG Lei(Key Laboratory of Genetic Breeding and Reproduction of Plateau and Mountain Animals of the Ministry of Education,Guizhou Key Laboratory of Animal Genetic Breeding and Reproduction,Guizhou University,Guizhou Guiyang 550025,China;College of Animal Science,Guizhou University,Guizhou Guiyang 550025,China)

机构地区：[1]贵州大学高原山地动物遗传育种与繁殖教育部重点实验室,贵州省动物遗传育种与繁殖重点实验室,贵州贵阳550025 [2]贵州大学动物科学学院,贵州贵阳550025

出　　处：《中国畜牧杂志》2022年第1期77-84,共8页Chinese Journal of Animal Science

基　　金：国家自然科学基金项目(31760652);贵州省肉羊产业技术体系建设项目。

摘　　要：为探究GPx3基因在产单、多羔黔北麻羊不同性腺组织中的表达规律及其蛋白质的结构与功能,本实验以黔北麻羊为研究对象,采集单、多羔黔北麻羊的下丘脑、垂体、子宫、输卵管和卵巢提取总RNA,运用qRT-PCR技术,分析GPx3基因mRNA在单、多羔黔北麻羊5个不同性腺组织中的表达水平;PCR法扩增克隆黔北麻羊GPx3基因的CDS区;生物信息学分析并预测黔北麻羊GPx3蛋白的结构与功能。qRT-PCR结果显示:GPx3基因在单、多羔黔北麻羊的5个不同性腺组织中均有表达,且均在卵巢的表达量最高;除下丘脑外,其余各组织在多羔组中的表达量均极限著高于单羔组。T克隆结果表明:黔北麻羊GPx3基因CDS区序列为681 bp,与NCBI上收录的山羊的mRNA序列同源性达99.85%。生物信息学分析发现:黔北麻羊GPx3蛋白可能位于细胞外,且具有信号肽,属于不稳定的亲水性分泌蛋白,其三级结构主要由无规则卷曲和α-螺旋构成;遗传进化树分析得出,黔北麻羊GPx3基因与绵羊的遗传距离最近,与鸡的遗传距离最远。本研究对GPx3基因进行了克隆和生物信息学分析,并揭示了GPx3基因在黔北麻羊不同性腺组织中的表达差异性,为进一步探究GPx3基因对黔北麻羊繁殖性状的调控机制提供基础。In order to explore the expression of GPx3 gene in different gonad tissues of single and multiple lamb of Qianbei ma goat and the structure and function of its protein.In this study,the total RNA of hypothalamus,pituitary,uterus,fallopian tube and ovary of single and multiple lamb of Qianbei ma goat was collected.The expression levels of GPx3 gene mRNA in five different gonad tissues of single and multiple lambs of Qianbei ma goat were analyzed by qRT-PCR technology.The PCR method was used to amplify and clone CDS region of GPx3 gene of Qianbei ma goat.The structure and function of GPx3 protein in Qianbei ma goat were analyzed and predicted by bioinformatics.The results of qRT-PCR showed that GPx3 gene was expressed in five different gonad tissues of single and multiple lamb of Qianbei ma goat,and the expression in ovary was the highest.Except for hypothalamus,the expression levels of other tissues in the multiple-lambing group were extremely higher than that in the single-lambing group.The results of T cloning showed that the CDS region sequence of GPx3 gene of Qianbei ma goat was 681 bp,and the homology with mRNA sequence of goat registered on NCBI was 99.85%.Bioinformatics analysis revealed that GPx3 protein of Qianbei ma goat might be located outside the cells and have signal peptides,whichbelongs to unstable hydrophilic secretory protein,and its tertiary structure is mainly composed of irregular curl andα-helix.The genetic tree analysis showed that the genetic distance of GPx3 gene from Qianbei ma goat was the closest to that of sheep,and the genetic distance was the farthest from that of chicken.In this study,GPx3 gene was cloned and analyzed by bioinformatics,and the expression differences of GPx3 gene in different gonad tissues of Qianbei ma goat were revealed,which provided the basis for further exploration of the regulatory mechanism of GPx3 gene on reproductive traits of Qianbei ma goat.

关 键 词：黔北麻羊 GPx3基因 克隆 生物信息学分析 荧光定量PCR 

分 类 号：S827.2[农业科学—畜牧学]