努比亚山羊PDK4基因克隆、序列分析及表达水平研究  

Study on cloning,sequence analysis and expression level of PDK4 gene in Nubian goats

作　　者：蔡沛燃 邹菊红 胡艳 张叁保 宋颖 蒋钦杨[1] CAI Peiran;ZOU Juhong;HU Yan;ZHANG Sanbao;SONG Ying;JIANG Qinyang(College of Animal Science and Technology,Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学动物科学技术学院,南宁530004

出　　处：《黑龙江畜牧兽医》2022年第9期55-60,141,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：广西重点研发计划项目(桂科AB18221067);崇左市科技计划项目(崇科FC2018006)。

摘　　要：为了探究努比亚山羊PDK4基因的脂质代谢途径,试验以40~48周龄努比亚山羊为研究对象,取其心脏、肾脏、肝脏、脾脏、背最长肌及皮下脂肪等组织/器官,分别提取总RNA,将其反转录为cDNA,采用PCR方法扩增PDK4基因片段,利用生物信息学软件分析PDK4基因序列,通过实时荧光定量PCR方法检测努比亚山羊不同组织/器官内PDK4基因的表达情况,同时以都安山羊为对照。结果表明:通过克隆成功获得努比亚山羊PDK4基因的CDS片段,其长度为1224 bp,共编码407个氨基酸残基;PDK4基因核苷酸序列与GenBank中的绵羊、牛、欧亚猪、马、人、小鼠的同源性分别为99.3%、97.6%、93.0%、92.5%、91.0%、85.0%;努比亚山羊PDK4蛋白氨基酸组成中亮氨酸含量较高,占总氨基酸数的10.8%,属于亲水性蛋白;努比亚山羊PDK4蛋白大部分是由α-螺旋和无规则卷曲组成,少部分由延伸链和β-转角组成。此外,努比亚山羊皮下脂肪中PDK4基因相对表达量最高,与其他组织比较差异显著(P<0.05),且心脏中PDK4基因相对表达量最低;努比亚山羊皮下脂肪中的PDK4基因相对表达量显著高于都安山羊(P<0.05)。说明PDK4基因可作为研究努比亚山羊脂肪沉积的主要候选基因之一。In order to explore the lipid metabolism pathway of PDK4 gene in Nubian goats,Nubian goats aged from 40 to 48 weeks were used as subjects.Total RNA was extracted from heart,kidney,liver,spleen,muscle,subcutaneous fat and other tissues and organs,and reversed into cDNA.The PDK4 gene was amplified by PCR.The PDK4 gene sequence was analyzed by bioinformatics website.The expression of PDK4 gene in different tissues and organs of Nubian goats was detected by real-time quantitative PCR,and Du’an goats were used as control group.The results showed that the CDS gene fragment of the Nubian goats PDK4 gene was successfully obtained by cloning,which was 1224 bp in length and encoded a total of 407 amino acid residues.The nucleotide sequences of PDK4 gene were 99.3%,97.6%,93.0%,92.5%,91.0%and 85.0%,respectively,with sheep,cattle,Eurasian pig,horse,human and mouse in GenBank.The amino acid composition of PDK4 protein of Nubian goats was high in leucine,accounting for 10.8%of the total amino acid number,which belonged to hydrophilic protein.Nubian goats PDK4 protein was mainly composed ofα-helix and random curl,and a small part was composed of extended chain andβ-turn.In addition,the relative expression of PDK4 gene was the highest in subcutaneous fat of Nubian goats,which was significantly different from other tissues(P<0.05),and the lowest in heart of Nubian goats.The relative expression level of PDK4 gene in subcutaneous fat of Nubian goats was significantly higher than that of Du’an goats(P<0.05).The results indicated that PDK4 gene could be one of the main candidate genes for studying fat deposition in Nubian goats.

关 键 词：努比亚山羊 都安山羊 PDK4基因 克隆 生物信息学 

分 类 号：S826[农业科学—畜牧学] Q78[农业科学—畜牧兽医]