鲁中肉羊TSHR多态性与产羔数的关联及生物信息学分析

Polymorphism and Bioinformatics Analysis of TSHR Gene and Its Association with Litter Size in Luzhong Mutton Sheep

本研究旨在探究TSHR基因g.89431097C>G位点多态性与鲁中肉羊产羔数之间的关系,并进行相关生物信息学分析,以期为TSHR基因编码蛋白功能机制研究提供基础数据并为鲁中肉羊高繁殖力分子育种提供新的遗传标记。利用全基因组重测序结合Sequenom MassARRAY?SNP技术对鲁中肉羊TSHR基因g.89431097C>G位点进行检测,并与其产羔数进行关联分析,利用Protparam等软件对TSHR基因进行生物信息学分析。分型结果表明:g.89431097C>G位点存在GG、GC和CC 3种基因型;群体遗传学分析表明,g.89431097C>G位点在鲁中肉羊中表现为中度多态(0.25<PIC<0.5);卡方适合性检验结果表明,该位点在鲁中肉羊中处于哈代温伯格平衡状态(p>0.05);关联分析表明,g.89431097C>G位点GG型第一胎、第二胎、第三胎产羔数均显著高于GC与CC型(p<0.05)。生物信息学分析表明绵羊TSHR基因编码764个氨基酸组成不稳定碱性疏水性蛋白,存在信号肽,存在7个跨膜域,预测存在6个N-糖基化位点、6个O-糖基化位点和85个磷酸化位点,预测存在二硫键,绵羊与山羊、家牛等12个物种TSHR基因编码区生物进化高度保守。

To elucidate the association between TSHR gene g.89431097 C>G polymorphism and litter size in Luzhong mutton sheep and conduct relevant bioinformatics analysis,in order to explore the genetic characters of this gene and the function mechanism of its encoding protein and provide new genetic markers for the molecular breeding of Luzhong mutton sheep with high fertility,Sequenom Mass ARRAY~?SNP assay was applied to genotype g.89431097 C>G locus of TSHR gene and the bioinformatics analysis was performed.The result of genotyping showed that three genotypes(GG,GC and CC)were found at g.89431097 C>G locus in Luzhong mutton sheep.Population genetic analysis showed that g.89431097 C>G locus was at moderate polymorphism(0.25<PIC<0.5)in Luzhong mutton sheep.Theχ~2 test indicated that the g.89431097 C>G locus of Luzhong mutton sheep was under Hardy-Weinberg equilibrium(p>0.05).Association analysis indicated that litter size of Luzhong mutton sheep with GG genotype was significantly higher than that with GC and CC genotypes(p<0.05).Bioinformatics analysis revealed that the encoding protein of sheep TSHR is an unstable,alkaline and hydrophobicity fat-soluble protein with signal peptide,disulfide bond,7 transmembrane domains,6 potential N-glycosylation sites,6 potential O-glycosylation sites and 85 phosphorylation sites,the encoding protein of sheep TSHR is homologous between sheep and 12 species including goat and cattle.