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多浪羊青春期启动下丘脑基因启动子区域甲基化与mRNA表达分析

Analysis of Methylation and mRNA Expression of Gene Promoter Region in the Hypothalamus of Dolang Sheep During Pubertal Initiation.

作者信息

Sui Zhiyuan, Zhang Zhishuai, Zhang Yongjie, Zhang Jihu, Li Qingjin, Xing Feng

机构信息

Key Laboratory of Tarim Animal Husbandry Science and Technology, Xinjiang Production and Construction Group, College of Animal Science, Tarim University, Alaer, China.

出版信息

DNA Cell Biol. 2023 Mar;42(3):130-139. doi: 10.1089/dna.2022.0281. Epub 2023 Feb 21.

Abstract

plays an important role in puberty initiation in sheep. This study aimed to discuss the correlation between different growth periods and the methylation status of cytosine-guanine dinucleotide (CpG) islands in the promoter region of the gene in the Dolang sheep's hypothalamus. In this study, the sequence of the gene promoter region in Dolang sheep was obtained by cloning and sequencing, and methyl groups of the CpG island of the gene promoter in the hypothalamus were detected by bisulfite sequencing PCR during the three periods of prepuberty, adolescence, and postpuberty in Dolang sheep. expression in the Dolang sheep's hypothalamus was detected by fluorescence quantitative PCR at three stages: prepuberty, puberty, and postpuberty. In this experiment, the 2993-bp promoter region was obtained, and it was predicted that there was a CpG island containing 15 transcription factor binding sites and 12 CpG sites, which may play a role in gene expression regulation. Overall, methylation levels increased from prepuberty to postpuberty, while expression levels decreased, indicating that expression was negatively correlated with promoter methylation levels. Variance analysis showed significant differences in the methylation status of CpG5, CpG7, and CpG9 between pre- and postpuberty ( < 0.05). Our data show that expression is increased by demethylation of promoter CpG islands, with CpG5, CpG7, and CpG9 implicated as critical regulatory sites.

摘要

在绵羊青春期启动过程中发挥重要作用。本研究旨在探讨多浪羊下丘脑该基因启动子区域不同生长时期与胞嘧啶 - 鸟嘌呤二核苷酸(CpG)岛甲基化状态之间的相关性。在本研究中,通过克隆和测序获得多浪羊该基因启动子区域的序列,并在多浪羊青春期前、青春期和青春期后三个时期,采用亚硫酸氢盐测序PCR检测下丘脑该基因启动子CpG岛的甲基基团。通过荧光定量PCR在青春期前、青春期和青春期后三个阶段检测多浪羊下丘脑该基因的表达。在本实验中,获得了2993 bp的该基因启动子区域,预测其存在一个包含15个转录因子结合位点和12个CpG位点的CpG岛,可能在基因表达调控中发挥作用。总体而言,从青春期前到青春期后甲基化水平升高,而该基因表达水平降低,表明该基因表达与启动子甲基化水平呈负相关。方差分析显示青春期前和青春期后CpG5、CpG7和CpG9的甲基化状态存在显著差异(P < 0.05)。我们的数据表明,启动子CpG岛去甲基化可增加该基因表达,其中CpG5、CpG7和CpG9被认为是关键调控位点。

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