欧洲黑杨MARs的分离克隆及序列分析

为获得林木植物核基质附着区(MARs)序列,诱导欧洲黑杨(Populus nigra)愈伤组织并制备悬浮细胞,裂解释放细胞核,除去非MARs的游离DNA片段,得到核基质。碱性缓冲液分离残留DNA与结合蛋白,回收并克隆残留DNA,测序后得到两个具有MARs序列特征的DNA片段A7和A23。序列特征分析结果表明它们都含有MARs的特征基元,通过与其他已发表的MARs序列比较,认为是从杨树中分离到的两个MARs片段,并预测了其功能。该研究是首次从木本植物获得MARs序列。

Isolation and Sequence Analysis of MARs from Populus nigra

In order to obtain matrix attachment regions(MARs) from woody plants,calli of Populus nigra were induced from young leaves,and suspension cultured cells were prepared from the calli and used as starting materials for MARs isolation.The nuclei were released after digestion of the cells with pectinase and cellulase,and nuclear matrixes were obtained after removal of the non-MARs segments.Separation of residual DNA and proteins were further performed by employing a high saline buffer,and the residual DNA was recovered and cloned.Two DNA fragments named A7 and A23 with typical MARs characteristics were obtained.Sequence analysis showed that both of the two isolated DNA fragments have the cha-racteristic motifs of MARs.Comparison with other known MARs sequences suggested that these two fragments were two new MARs isolated from Populus,and their functions were also predicted.To our know-ledge,this is the first report of MARs isolated from a woody plant.