美仁牦牛PHB基因克隆及组织表达  

Cloning and Tissue Expression PHB Gene in Meiren Yak

作　　者：杨国武 王佟 马小勇 扎老 路建卫 赵雪 阎萍[2,3] 梁春年 YANG Guowu;WANG Tong;MA Xiaoyong;ZHA Lao;LU Jianwei;ZHAO Xue;YAN Ping;LIANG Chunnian(Life Science and Engineering College,Northwest Minzu University,Lanzhou 730124,China;Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences,Key Laboratory of Yak Breeding Engineering of Gansu Province,Lanzhou 730050,China;Key Laboratory of Animal Genetics and Breeding on the Tibetan Plateau,Ministry of Agriculture and Rural Affairs,Lanzhou 730050,China;Animal Husbandry Station of Zogaidoma Township,Hezuo 747000,China;Quality and Safety Inspection Center of Agricultural and Livestock Products in Hezuo,Hezuo 747000,China)

机构地区：[1]西北民族大学生命科学与工程学院,甘肃兰州730124 [2]中国农业科学院兰州畜牧与兽药研究所,甘肃省牦牛繁育工程重点实验室,甘肃兰州730050 [3]农业农村部青藏高原畜禽遗传育种重点实验室,甘肃兰州730050 [4]合作市佐盖多玛乡畜牧站,甘肃合作747000 [5]合作市农畜产品质量安全检测检验中心,甘肃合作747000

出　　处：《华北农学报》2023年第5期218-226,共9页Acta Agriculturae Boreali-Sinica

基　　金：国家重点研发计划(2021YFD1600200);甘肃省基础研究创新群体项目(20JR5RA580);合作市牦牛种质提升与提质增效项目、牦牛现代肉牛牦牛产业技术体系(CARS-37);中国农业科学院创新工程(25-LIHPS-01);甘肃省科技重大专项(21ZD10NA001;GZGG-2021-1)。

摘　　要：为研究美仁牦牛抗增殖蛋白(PHB)的结构及功能,并探究其在7日龄牛和成年牛的各组织中的表达情况。以美仁牦牛肾脏组织cDNA为模板克隆美仁牦牛PHB基因序列。对基因序列进行生物信息学分析以及对蛋白质进行理化性质预测,实时荧光定量PCR(RT-qPCR)技术检测PHB基因在美仁牦牛2个年龄段8个组织中的相对表达量。结果表明,美仁牦牛PHB基因的CDS区全长819 bp,共编码272个氨基酸;同源性比对发现美仁牦牛与普通牛的同源性最高(99.1%);美仁牦牛PHB蛋白分析显示,PHB蛋白内存在1个N-糖基化潜在位点和2个O-糖基化潜在位点,其氨基酸等电点为5.55,不稳定系数(Ⅱ)为46.07,总平均疏水性为0.045,预测为不稳定疏水性蛋白质;亚细胞定位显示,PHB蛋白位于线粒体(30.4%),蛋白内共有潜在磷酸化位点15个,PHB蛋白高级结构由α-螺旋(56.25%)、无规则卷曲(22.43%)、延伸链(16.54%)和β-转角(4.78%)相互连接而成。RT-qPCR结果显示,美仁牦牛2个发育阶段的8个组织中均有PHB基因的表达。在对比2个时期的PHB基因组织表达规律后发现,7个组织的PHB基因表达量随美仁牦牛的生长发育而降低,在美仁牦牛肝脏、脾脏、肺脏、脂肪和睾丸组织中,PHB基因表达量随年龄显著降低(P<0.05)。In order to study the structure and function of PHB gene in Meiren yak,and to explore its expression in different tissues at 7-day-old and adult cattle.This study cloned the coding region of PHB gene from Meiren yak kidney tissue cDNA as a template.The gene sequence was analyzed by bioinformatics and the physical and chemical properties of the protein were predicted.The relative expression levels of PHB gene in 8 tissues of Meiren yak at two ages were detected by Real-time Fluorescence Quantitative(RT-qPCR).The results showed that the CDS region of the PHB gene of Meiren yak was 819 bp in length,encoding 272 amino acids.The results of homology analysis showed that Meiren yak had the highest homology with common cattle(99.1%).The analysis of PHB protein showed that there was one potential N-glycosylation site and two potential O-glycosylation sites in the PHB protein.Its amino acid isoelectric point was 5.55,the instability coefficient(Ⅱ)was 46.07,and the total average hydrophobicity was 0.045,which was predicted to be an unstable hydrophobic protein.Subcellular localization showed that PHB protein was located in the mitochondria(30.4%),and there were 15 potential phosphorylation sites in the protein.The high-level structure of PHB protein was interconnected byα-helix(56.25%),random coil(22.43%),extended chain(16.54%)andβ-corner(4.78%).The results of RT-qPCR showed that the PHB gene was expressed in eight tissues of Meiren yak at two developmental stages.The expression of PHB gene in 7 tissues decreased with the growth and development of Meiren yak.The expression of PHB gene in liver,spleen,lung,fat and testis decreased significantly with age(P<0.05).

关 键 词：美仁牦牛 PHB基因 克隆 生物信息学 表达谱 

分 类 号：S823[农业科学—畜牧学] Q78[农业科学—畜牧兽医]