长白猪×蓝塘猪及其杂种基因组甲基化片段克隆与分析

Cloning and Analysis of DNA Methylation Fragment in Landrace× Lantang Pigs and Their Hybrids

应用甲基化敏感扩增多态性(MSAP)技术对6头长白公猪和50头蓝塘母猪及51头长白×蓝塘杂交F1代3个群体基因组DNA胞嘧啶甲基化位点进行检测,旨在克隆和分析父母代猪及其杂种F1代之间基因组DNA共同甲基化片段和差异片段,并找到其同源基因。结果显示,从MSAP条带中分离、克隆得到18条3个群体共同的甲基化片段、10条母代独有的甲基化片段和9条杂交一代独有的甲基化片段,其中有1条3个群体共有的甲基化片段通过EST拼接和电子延伸后在NCBI数据库中找到同源基因,即猪类酪氨酸蛋白激酶Lyn基因(GeneID:LOC100152890,序列号:XM_001926250)。结果表明,长蓝杂交F1代与其父母代之间的基因组甲基化存在异同,为通过MSAP技术克隆猪基因组DNA甲基化片段及寻找其对应的甲基化基因提供可能,也会为将来研究这些甲基化基因表达调控机制奠定基础。

To clone and analyze common and different methylation fragments between parents pigs and their F1 hybrids,and find their homologous genes,genome DNA cytosine methylation was detected in 6 Landrace boars,50 Guangdong Lantang sows and 51 Landrace × Lantang F1 hybrids by MSAP in this study.The results showed 18 co-methylation fragments in three populations,10 only mothers＇ methylation fragments and 9 only F1 hybrids＇ methylation fragments were separated and cloned from MSAP fragments,among all of these fragments,one which resided in three populations was found a homologous gene in NCBI database after spliced with EST and electronic extension,i.e.,Sus scrofa similar to Tyrosine protein kinase Lyn gene（GeneID：LOC100152890,SN：XM_001926250）.The results indicated the DNA methylation had some differences between Landrace × Lantang F1 hybrids and their parents.This study provided the possibility in cloning pig genome methylation fragments and seeking their homologous genes by MSAP,and also could lay the basis for a future research on methylation gene expression and regulation.