多浪羊TRHR基因克隆及不同初情期阶段组织差异性表达分析OA

The aim of this study was to clone the TRHR(Thyrotropin-releasing hormone receptor)gene of Duolang sheep and to detect the expression of the TRHR gene in the tissues related to reproduction,so as to provide a theoretical basis for exploring the role of the TRHR gene in sheep reproduction.The cDNA of puberty pituitary gland of Duolang sheep was used as template for the experiment,and the TRHR gene was amplified by RT-PCR and the resulting product was cloned and sequenced.Bioinformatics software was used to predict the structure and physicochemical properties of TRHR protein in Duolang sheep.The expression levels of TRHR gene in five tissues related to reproduction in three pe-riods before and after puberty in Duolang sheep were also detected by qPCR.The results showed that the sequence size of the cloned TRHR gene was 1 373 bp,of which the CDS region was 1 197 bp,with 99.83％similarity to the mRNA se-quence of sheep predicted on GenBank.Phylogenetic tree analysis revealed that the TRHR gene of Duolang sheep had the closest genetic distance to Oryx dammah and the furthest to Lontra canadensis.qPCR assay showed that the TRHR gene of Duolang sheep was significantly expressed in different periods and tissues,and at the puberty stage,the expres-sion of the TRHR gene in the pubertal pituitary gland was significantly higher than that in other tissues(P＜0.05).In sumamry,the present study successfully cloned the sequence of the coding region of TRHR gene in Duolang sheep and found that the gene showed high expression characteristics in the pubertal pituitary gland of Duolang sheep.