牦牛DJ-1基因克隆、生物信息学及组织表达分析  

Cloning,Bioinformatics,and Tissue Expression Analysis of Yak DJ-1 Gene

作　　者：申金伟 路建卫 赵雪 扎老 成述儒[3] 梁春年[1,2] SHEN Jinwei;LU Jianwei;ZHAO Xue;ZHA Lao;CHENG Shuru;LIANG Chunnian(Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences,Key Laboratory of Yak Breeding Engineering 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;Gansu Agricultural University,Lanzhou 730124,China;Animal Husbandry and Veterinary Station,Zuogaimanma Town,Hezuo 747000,China;Agricultural and Animal Products Quality Safety Inspection Center,Hezuo 747000,China)

机构地区：[1]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 [2]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 [3]甘肃农业大学,甘肃兰州730124 [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》2024年第2期192-199,共8页Acta Agriculturae Boreali-Sinica

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

摘　　要：为了研究牦牛DJ-1基因的结构及功能,为后续深入研究牦牛DJ-1基因的生物学功能提供了重要的理论依据。以美仁牦牛脂肪组织cDNA为模板,利用RT-PCR克隆牦牛DJ-1基因的编码区序列(CDS),并且利用基因序列进行生物信息学分析,预测结构域及蛋白质理化性质等,再利用RT-qPCR技术进行组织表达分析。结果表明,美仁牦牛DJ-1基因的CDS区全长570 bp,共编码189个氨基酸;牦牛DJ-1结构域预测显示,在DJ-1氨基酸序列的29—168位中有1个含有Pfam的PfpI结构域;通过对DJ-1蛋白结构和功能进行预测,结果显示,无跨膜结构且无信号肽区域,分子质量20.03531 ku,原子总数2870,理论等电点6.84,不稳定系数28.37,表示为稳定的蛋白质;脂肪系数101.11,总平均亲水系数-0.004,为亲水蛋白,共有11个磷酸化位点;同时,亚细胞定位预测结果表明,美仁牦牛DJ-1蛋白主要分布于细胞质和线粒体中,系统进化树表明,美仁牦牛与野牦牛和欧洲牛亲缘关系最近,与北极狐和宽吻海牛亲缘关系最远;RT-qPCR结果表明,在成年美仁牦牛的心脏、肝脏、脾脏、肺脏、肌肉、脂肪和睾丸组织中均有所表达,在心脏组织和肌肉组织中表达水平最高,在肝脏和睾丸组织中表达水平最低。In order to study the structure and function of DJ-1 gene in yaks,this experiment used Meiren yak adipose tissue cDNA as a template,cloned the coding sequence(CDS)of yak DJ-1 gene using RT-PCR,and performed bioinformatics analysis,including domain prediction and protein physicochemical properties prediction of the gene sequence.Then,tissue expression analysis was performed using RT-qPCR.The results showed that the CDS region of the DJ-1 gene in Meiren yak had a total length of 570 bp and encoded 189 amino acids;the prediction of the DJ-1 domain in yaks showed that there was one PfpI domain containing the Pfam in positions 29-168 of the DJ-1 amino acid sequence.By predicting structure and function of the DJ-1 protein,the results showed no transmembrane structure and no signal peptide region;molecular weight was 20.03531 ku,total number of atoms was 2870,theoretical isoelectric point was 6.84,instability coefficient was 28.37,and expressed as a stable protein;the fat coefficient was 101.11,with a total average hydrophilicity coefficient of-0.004.It was a hydrophilic protein with a total of 11 phosphorylation sites.Meanwhile,the results of subcellular localization prediction indicated that the DJ-1 protein of Meiren yak was mainly distributed in the cytoplasm and mitochondria.The phylogenetic tree showed that Meiren yak had the closest genetic relationship with wild yaks and European cattle,and the farthest genetic relationship with Arctic foxes and wide snouted manatees;the RT-qPCR results showed that it was expressed in the heart,liver,spleen,lungs,muscle,fat,and testicular tissues of adult Meiren yaks,with the highest expression level in the heart and muscle tissues,and the lowest expression level in the liver and testicular tissues.

关 键 词：美仁牦牛 DJ-1基因 生物信息学分析 系统进化 

分 类 号：S823.8[农业科学—畜牧学] Q78[农业科学—畜牧兽医]