鸡、鹌鹑及其杂交种早期胚胎bcl-2基因甲基化的分析

采用甲基化特异性PCR(MSP)方法进行鸡、鹌鹑及其属间杂交种早期胚胎发育期60、66、72、84、96、108、120 h 7个不同胚龄胚胎组织bcl-2基因启动子区CpG岛甲基化状态的对比分析,探讨bcl-2基因甲基化对鸡与鹌鹑属间杂交种早期胚胎发育的影响。结果显示,正常发育的鸡胚和鹌鹑胚龄在60、66、72、120 h均呈高甲基化状态,84和96 h呈非甲基化状态,而鸡与鹌鹑属间杂交种胚胎在60、66、72、96、108和120 h则与鸡、鹌鹑不同,呈现出甲基化或非甲基化无规律性并存,甚至检测不到甲基化状态;84 h则只检测到非甲基化状态。鸡与鹌鹑杂交种早期胚胎组织中bcl-2 基因启动子区CpG岛的异常甲基化有可能是引起鸡与鹌鹑属间杂交种胚胎早期死亡的一个重要影响因素。

Analysis of bcl-2 Gene Methylation in Early Embryonic of Chicken,Quail and its Hybrid

This paper was conducted to study the methylation of bcl-2 gene and its death relevance with the hybrids of chicken-quail in early embryonic stage.The CpG island methylation status of bcl-2 gene promoter region was detected by the methylation-specific PCR in seven different chicken quail and its hybrid fetation stages(60,66,72,84,96,108 and 120 h).The results showed that the hypermethylation(60,66,72 and 120 h) and unmethylation(84 h,96 h) was detected in the chicken and quail embryos;the hypermethylation of bcl-2 gene promoter CpG islands was detected in the chicken and quail embryos treated by M.sssⅠ enzyme.Compared with the chicken and quail,the abnormal methylation appears in chicken-quail hybrid embryos,and methylation or unmethylation was detected in some sample(60,66,72,96,108 and 120 h),undetected in the others.However,unmethylation was tested in 84 h hybrid embryos.An important influence factor in early death of chicken-quail hybrid embryos were probably CpG island abnormal methylation of bcl-2 gene promoter region.