PEG11基因在体细胞核移植牛中印迹和DNA甲基化状态分析  

Genomic imprinting and DNA methylation status of PEG11 in somatic cell nuclear transfer cattle

作　　者：张明月[1] 杨文志[1] 石运娇[1] 张萃[1] 陈玮娜[2] 崔亚丽[1] 张巍巍[1] 李宁[3] 李世杰[1] ZHANG Ming-yue YANG Wen-zhi SHI Yun-jiao ZHANG Cui CHEN Wei-na CUI Ya-liI ZHANG Wei-wei LI Ninga LI Shi-jie(College of Life Science, Hebei Agricultural Uni- versity, Baoding, Hebei 071001, China College of Medical Science, Hebei University, Baoding, Hebei 071001,China The State Key Laboratory for Agrobiotechnology in Livestock and Poul- try, China Agricultural University, Beij ing 100083, China)

机构地区：[1]河北农业大学生命科学学院,河北保定071001 [2]河北大学药学院,河北保定071001 [3]中国农业大学畜禽农业生物技术国家重点实验室,北京100083

出　　处：《中国兽医学报》2016年第12期2154-2159,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31372313);河北省自然科学基金资助项目(C2011204001)

摘　　要：为了分析PEG11基因在体细胞核移植(somatic cell nuclear transfer,SCNT)牛中的印记以及重编程状态,本研究应用RT-PCR产物直接测序法对PEG11基因在自然繁殖牛和新生死亡SCNT牛7个组织(心、肝、脾、肺、肾、肌肉和脂肪)中的表达进行了分析。结果发现PEG11基因在自然繁殖牛的7个组织中均为单等位基因表达,表明PEG11基因在牛中是印记的;在SCNT牛肺脏中PEG11基因为双等位基因表达,而在其余6个组织中为单等位基因表达。进而用亚硫酸氢盐测序法分析自然繁殖牛和SCNT牛肺脏PEG11基因中54CpGs位点的甲基化状态,发现PEG11基因在SCNT牛肺脏中呈现与自然繁殖牛相似的高甲基化状态(97.26%),推测PEG11基因在新生死亡SCNT牛肺脏中印记紊乱可能是导致SCNT牛肺脏发育缺陷的原因之一,PEG11基因内部CGIs的甲基化不参与调控PEG11基因的基因组印记。Somatic cell nuclear transfer（SCNT） is an inefficient process,and the incomplete or aberrant reprogramming of donor nuclear is consided as the main reason. Genomic imprinting and DNA methylation play important roles in mammalian development,and abnormal expression or aberrant DNA methylation status of imprinted genes have been frequently observed in SCNT animals. PEG11 is a paternally expressed imprinted gene identified in human,and its imprinting status has not been established in cattle. In an effort to determine the reprogramming of PEG11 in SCNT cattle,we firstly analyzed the imprinting status of PEG11 in seven organs （heart, liver, spleen, kid- ney, lung, subcutaneous fat and skeletal muscles） of naturally breed calves by directly sequencing of the RT-PCR products,and monoallelic expressions of PEGll were observed in all detected tis- sues, suggested that PEGll is imprinted in cattle. However,in neonatal died SCNT calves, abnor- mal biallelic expressions of PEGll were found in lung tissues. Then the DNA methylation statuses of 54 CpGs position in PEG11 were analyzed in lung tissues of SCNT cattle. PEG11 exhibited seriously hypermethylated vines. Our results suggest the defects of lung organs death of SCNT animals. （97. 26%） in SCNT cattle, which was similar to naturally breed bo- that the aberrant biallelic expression of PEG11 may be associated with in died SCNT bovines, which is one of the main reasons for neonatal

关 键 词：PEG11基因 印记状态 体细胞核移植牛 DNA甲基化 器官缺损 

分 类 号：S814.8[农业科学—畜牧学]