畜牧兽医学报2017，Vol.48Issue(12)：2277-2285,9.DOI:10.11843/j.issn.0366-6964.2017.12.007
克隆山羊成纤维细胞IGF2-H19基因座甲基化分析
IGF2-H19 Locus Methylation Status in Cloned Goat Fibroblast Cells
摘要
Abstract
The aim of this experiment was to study the influence of somatic cell nuclear transfer on IGF2-H19 locus methylation in cloned goat fibroblast cells.Goat ear fibroblast cells (GFC,control group) and cloned goat ear fibroblast cells (CFC,experimental group) were used as experimental materials.When cells in the 2 groups were cultured to passage 5,using cell counting method for cell growth curves,flow cytometer for cell apoptosis,real-time PCR for gene relative expression level and bisulfite sequencing PCR for the methylation status of differentially methylated region (DMR).The results showed that cell growth curves in the 2 groups were typical "S"-shaped,but apoptosis rate in CFC group was increased significantly(P＜0.01) compared with that in GFC group.The expression level of Dnmt1 (P＜0.01),Dnmt3b (P＜0.01),Tet1 (P＜0.05),Tet2 (P＜0.05),H19 (P＜0.05) and IGF2 (P＜0.01) in CFC group were decreased significantly compared with that in GFC group.However,no statistical differences were found at the expression levels of Tet3,Dnmt3a and IGF2R between the 2 groups.Compared with GFC group,the methylation level of IGF2 DMR1 (74.1％ vs.57.8％,P＜0.01) and DMR2 (76.8％ vs.40.0％,P＜0.01) were decreased significantly,while the methylation level of IGF2-H19 imprinting control region (ICR) was increased significantly (68.8％ vs.84.0％,P＜0.01) in CFC group.Somatic cell nuclear transfer resulted in aberrant methylation of IGF2-H19 locus by influencing Dnmts and Tets genes expression,which could affect genomic imprinting,and then influence the efficiency of serial recloning.关键词
体细胞核移植/再克隆/IGF2-H19/DNA甲基化/山羊耳成纤维细胞Key words
somatic cell nuclear transfer/recloning/IGF2-H19/DNA methylation/goat ear fibroblast cells分类
农业科技引用本文复制引用
刘孜斐,邓明田,任才芳,万永杰,王锋..克隆山羊成纤维细胞IGF2-H19基因座甲基化分析[J].畜牧兽医学报,2017,48(12):2277-2285,9.基金项目
国家自然科学基金(31301973) （31301973）
