绵羊FBXL3基因表达及其多态性与季节性发情的相关性分析  

FBXL3 Gene Expression and Correlation Analysis Between Its Polymorphism and Seasonal Estrus in Sheep(Ovis aries)

作　　者：杨阳 钟英杰 姜雨[3] 储明星[1] 刘秋月 YANG Yang;ZHONG Ying-Jie;JIANG Yu;CHU Ming-Xing;LIU Qiu-Yue(Key Laboratory of Animal Genetics,Breeding and Reproduction of Agriculture and Rural Affairs/Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Institute of Genetics and Developmental Biology/Innovation Academy for Seed Design,Chinese Academy of Sciences,Beijing 100101,China;Key Laboratory of Animal Genetics,Breeding and Reproduction of Shaanxi Province/College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所/农业农村部动物遗传育种与繁殖重点实验室,北京100193 [2]中国科学院遗传与发育生物学研究所/种子创新研究院,北京100101 [3]西北农林科技大学动物科技学院/陕西省动物遗传育种与繁殖重点实验室,杨凌712100

出　　处：《农业生物技术学报》2022年第1期75-84,共10页Journal of Agricultural Biotechnology

基　　金：国家转基因生物新品种培育科技重大专项(2016ZX08009-003-006);国家肉羊产业技术体系专项(CARS-38);中国农业科学院科技创新工程(ASTIP-IAS13)。

摘　　要：季节性发情限制了绵羊(Ovis aries)的繁殖效率,因此,探究影响绵羊季节性发情的关键基因具有重要意义。为探究通过全基因组重测序(whole genome resequencing, WGRS)筛选出的绵羊F-box富含亮氨酸重复蛋白3 (F-box and leucine-rich repeat protein 3, FBXL3)基因表达水平及其多态性与绵羊季节性发情之间的关系,本研究选取苏尼特羊为对象,利用qPCR方法分析FBXL3基因在不同组织中的表达及SNP对其表达的影响;利用Sequenom MassARRAY;SNP技术对2种发情模式的绵羊(常年发情组和季节性发情组) FBXL3基因g.52551820G>A位点多态性进行检测,并与绵羊的季节性发情性状进行关联分析;同时,利用生物信息学方法分析g.52551820G>A位点突变对mRNA二级结构和蛋白质性质的影响。结果表明,FBXL3基因在苏尼特羊多种组织中广泛表达;FBXL3基因g.52551820G>A位点存在3种基因型GG、GA和AA,在肝脏(P<0.01)、肾脏(P<0.01)和肾上腺(P<0.05)中野生型GG表达量显著高于突变型AA;在季节性发情品种中G为优势等位基因,在常年发情品种中A为优势等位基因,该位点的基因型频率及等位基因频率在季节性发情和常年发情品种中都达到了差异极显著水平(P<0.01);群体遗传学分析结果显示,FBXL3基因g.52551820G>A位点在草原型藏羊和杜泊羊中表现为低度多态(PIC<0.25),在滩羊、苏尼特羊、小尾寒羊和萨福克羊中表现为中度多态(0.25A位点突变可能是绵羊季节性发情的潜在分子标记,本研究为季节性发情的分子机制研究提供基础资料。Seasonal estrus limits reproduction efficiency of sheep(Ovis aries), therefore, it is of great significance to explore key genes that affect the seasonal estrus of sheep. In order to explore the expression level and polymorphism of the F-box and leucine-rich repeat protein 3(FBXL3) gene which was found by whole genome resequencing(WGRS) and its association with seasonal estrus in sheep(Ovis aries), the present study selected Sunite sheep as research object, qPCR was used to investigate the expression of FBXL3 gene in different tissues and the influence of SNP on its expression. SequenomMassARRAY;SNP was applied to detect polymorphism of g.52551820G>A locus of FBXL3 gene in 2 different estrus patterns(year-round estrus group and seasonal estrus group). Then correlation analysis was carried out with seasonal estrus traits of sheep. Meanwhile, bioinformatics methods were used to analyze the effects of mutation on mRNA secondary structure and protein properties. The results showed that FBXL3 gene was widely expressed in the tissues of Sunite sheep, and 3 genetypes of GG, GA, and AA existed in the locus of FBXL3 g. 52551820G>A. The expression of GG genotype was significantly higher than AA genotype in liver(P<0.01), kidney(P<0.01),and adrenal gland(P<0.05). The G was the dominant allele in seasonal estrus breed, while A was the dominant allele in year-round estrus breed. The genotype frequency and allele frequency of this locus were extremely significant different between seasonal estrous and year-round estrous breeds(P<0.01). Population genetic analysis showed that the g.52551820G>A locus of FBXL3 gene was at low polymorphism(PIC<0.25)in Prairie Tibetan sheep and Dorper sheep, and was at moderate polymorphism(0.25<PIC<0.50) in Tan sheep, Sunite sheep, Small Tail Han sheep and Suffolk sheep. Moreover, bioinformatics analysis showed that the hydrophilicity of FBXL3 amino acid residues near the mutation site decreased slightly, and the phosphorylation of the protein also changed. Altogether, the mutation at g.52551820G>

关 键 词：绵羊 季节性发情 F-box富含亮氨酸重复蛋白3基因(FBXL3) 组织表达 生物信息学分析 

分 类 号：S826[农业科学—畜牧学]