PROSER2基因在牛中的印记表达和DNA甲基化分析  

The study of imprinted expression and DNA methylation of PROSER2 gene in cattle

作　　者：张银蛟 杨利丹 郑云畅 李树静 余文莉 陈玮娜 李世杰[1] ZHANG Yinjiao;YANG Lidan;ZHENG Yunchang;LI Shujing;YU Weni;CHEN Weina;LI Shijie(College of Life Science,Hebei Agricultural University,Baoding 071001,China;Hebei Provincial Dairy Cow Breeding Engineering Technology Research Center,Shijiazhuang 050000,China;Hebei Cattle Industry Technology Research Institute,Shijiazhuang 050000,China;College of Medical Science,Hebei University,Baoding 071000,China)

机构地区：[1]河北农业大学生命科学学院,河北保定071000 [2]河北省奶牛良种繁育技术创新中心,河北石家庄050000 [3]河北省牛产业技术研究院,河北石家庄050000 [4]河北大学中医学院,河北保定071000

出　　处：《河北农业大学学报》2024年第3期85-92,共8页Journal of Hebei Agricultural University

基　　金：河北省自然科学基金(C2023204145,C2022201058);河北省现代农业产业技术体系建设专项资金(HBCT2024240206,HBCT2024240101).

摘　　要：为了研究牛PROSER2基因的印记状态和表观遗传修饰机制,本研究首先应用荧光定量RT-PCR方法分析PROSER2基因在牛组织和胎盘中的表达,进而应用基于单核苷酸多态性(SNP)的RT-PCR产物直接测序法分析等位基因表达状态,最后采用亚硫酸盐直接测序法对位于PROSER2基因启动子及第1个外显子区域和位于外显子4区域的2个CpG岛的甲基化状态进行了分析。结果显示,PROSER2基因在所有检测的6个组织(心脏、肝脏、脾脏、肺脏、肾脏、大脑)及胎盘中均广泛表达。PROSER2基因在牛的心脏、肝脏、脾脏、肺脏、肾脏、大脑以及胎盘中均为单等位基因表达。根据杂合胎盘父母的基因型,发现PROSER2基因在牛中是父源印记基因,这与其在人中的印记状态一致。对PROSER2基因启动子和第一个外显子处CpG岛区域的甲基化状态进行分析,在牛的心脏、肝脏、脾脏、肺脏、肾脏、大脑和胎盘中均发现1个差异甲基化区,说明DNA的甲基化修饰可能参与PROSER2基因在荷斯坦奶牛中的印记表达。本研究结果可为进一步研究PROSER2基因的功能和印记调控机制提供参考依据。The aim of this study was to investigate the imprinting status and epigenetic modification mechanism of bovine PROSER2 gene.The expression of PROSER2 gene was first analyzed in bovine tissues and placenta using quantitative RT-PCR.The direct sequencing of RT-PCR products were analyzed based-on single nucleotide polymorphism(SNP)for the imprinting status of PROSER2 gene.Finally,the methylation status of two CpG islands covered the promoter/exon 1 region and exon 4 regions of PROSER2 gene,was analyzed using bisulfite sequencing.The results showed that the PROSER2 gene was expressed in all the tested tissues including heart,liver,spleen,lung,kidney,brain and placenta.The PROSER2 gene was monoallelally expressed in the bovine heart,liver,spleen,lung,kidney,brain and placenta.PROSER2 gene was a paternally imprinted gene in cattle,which is consistent with its imprinting status in human.A differentially methylated region was found in the CpG island located in the promoter and the exon 1 of the PROSER2 gene in the bovine heart,liver,spleen,lung,kidney,brain and placenta.This suggests suggested that methylation modifications of DNA may be involved in imprinted expression of bovine PROSER2 gene.This study can provide a reference for the further study of the function and imprinting regulation mechanism of bovine PROSER2 gene.

关 键 词：DNA甲基化 基因组印记 牛 PROSER2基因 表观遗传 

分 类 号：S823[农业科学—畜牧学] Q78[农业科学—畜牧兽医]