牛ANO1基因为胎盘特异的父源印记基因  

Bovine(Bos taurus) ANO1 Gene is a Placental-specific Paternal Imprinting Gene

作　　者：谷书凯 李俊良 陈玮娜 张萃[1] 徐达[1] 李冬杰[3] 李世杰[1] GU Shu-Kai;LI Jun-Liang;CHENWei-Na;ZHANG Cui;XU Da;LI Dong-Jie;LI Shi-Jie(College of Life Science,Hebei Agricultural University,Baoding 071001,China;Department of Traditional Chinese Medicine,Hebei University,Baoding 071001,China;College of Bioscience and Bioengineering,Hebei University of Science and Technology,Shijiazhuang 050018,China)

机构地区：[1]河北农业大学生命科学学院,保定071001 [2]河北大学中医系,保定071001 [3]河北科技大学生物科学与工程学院,石家庄050018

出　　处：《农业生物技术学报》2019年第11期1996-2003,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31372312);河北省自然科学基金(No.C2016204092)

摘　　要：基因组印记(genomic imprinting)是一种表观遗传(epigenetics)现象,是指哺乳动物中部分基因表现出亲本等位基因表达的差异性,即只表达一方的亲本等位基因。ANO1(anoctamin 1)蛋白是钙激活氯通道的一个亚基,ANO1基因的过度表达与人类(Homo sapiens)多种癌症相关。该基因在人的胚胎和成年组织中为双等位基因表达,但在胎盘中为母源等位基因表达的印记基因。而ANO1基因在牛(Bos taurus)中的印记状态以及调控机理尚未见研究报道。为了分析ANO1基因在牛中的印记状态,首先通过PCR产物直接测序法在牛ANO1基因上找到SNP位点,进而对杂合子牛的组织和胎盘中的RNA进行反转录PCR(reverse transcription-PCR,RT-PCR),经产物直接测序发现ANO1基因在牛中的印记状态为胎盘特异性印记,且为母源等位基因特异性表达。进一步采用亚硫酸氢盐测序法分析牛胎盘、牛精子以及成年牛组织中ANO1基因启动子区等位基因特异的甲基化状态,发现ANO1基因启动子甲基化不参与该基因的基因组印记表达。本研究结果表明牛ANO1基因为胎盘特异的父源印记基因,可为进一步研究ANO1基因表达与相关疾病的关联提供参考依据。In mammals,genomic imprinting is an epigenetic phenomenon by which certain genes are expressed from one allele,in a parent-of-origin-specific manner.ANO1(Anoctamin 1)protein is a subunit of calcium-activated chloride channel.Overexpression of ANO1 gene is associated with a variety of human(Homo sapiens)cancers.In human,ANO1 gene exhibits biallelic expression in embryos and adult tissues,but shows maternal allele expression in placenta.At present,the imprinting status and regulation mechanism of ANO1 gene in bovine(Bos taurus)have not been reported.In this study,in order to analyze the imprinting status of ANO1 gene in cattle,SNP was first found on ANO1 gene by direct sequencing of PCR products,and then reverse transcription-PCR(RT-PCR)was performed on RNA derived from tissues and placentas of heterozygous cattle.Direct sequencing showed that the imprinting status of ANO1 gene in cattle was placenta-specific imprinting and maternal allele-specific expression.The specific methylation status of the promoter region of ANO1 gene in bovine placenta,bovine sperm and adult bovine tissues was further analyzed by sulfite sequencing.It was found that the promoter methylation of ANO1 gene did not participate in the gene imprinting expression.The results showed that bovine ANO1 gene was a placenta-specific paternal imprinting gene,which provides a reference for further study of the relationship between ANO1 gene expression and related diseases.

关 键 词：牛 Anoctamin 1基因(ANO1) 印记状态 胎盘 DNA甲基化 

分 类 号：S823[农业科学—畜牧学] S813.3[农业科学—畜牧兽医]