FBXL3 Gene Expression and Correlation Analysis Between Its Polymorphism and Seasonal Estrus in Sheep (Ovis aries)
YANG Yang1, ZHONG Ying-Jie1, JIANG Yu3, CHU Ming-Xing1,*, LIU Qiu-Yue2,*
1 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; 2 Institute of Genetics and Developmental Biology/Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China; 3 Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province/College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China
Abstract：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>A locus of FBXL3 gene might be a potential molecular marker of seasonal estrus in sheep, which provides a basic reference for studying the molecular mechanism of seasonal estrus.
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