一个低氮诱导表达的水稻Dof转录因子OsDof-13的分离和转化

Isolation and Transformation of OsDof-13, a Member of Rice Dof Transcription Factor Family, Induced by Low Nitrogen Stress

通过生物信息学同源分析,在水稻cDNA数据库中检索到水稻Dof家族中的OsDof-13(AK119803)基因与玉米MNB1a转录因子具有很高的同源性。NCBI数据库注释,该基因编码202个氨基酸。依据Dof转录因子共有的特征,N端51～102AA可能为该基因的Dof结构域。Northern分析结果表明Os-Dof-13受低氮诱导后有明显的表达变化。亚细胞定位分析结果表明OsDof-13蛋白定位于细胞核。构建了OsDof-13的超表达载体,利用农杆菌介导法将其转化粳稻品种合江19,获得12个独立转基因植株,South-ern杂交结果显示OsDof-13基因的插入拷贝数分别为1～4个。

Bioinformatic analysis shows that OsDof-13 （AK119803）, a member office Dof family, is highly homologous to the Dof transcfiption factor MNBla in maize （Zea Mays）. In NCBI database OsDof-13 protein consists of 202 amino acids. According to the common character of the Dof transcription factors, the region of 51-102 amino acids at N-terminator of OsDof-13 protein is predicted to be a conserved Dof motif. Northern blot analysis verified that OsDof-13 was strongly induced by low nitrogen stress, while OsDof-13 protein was located in nucleus by sub-cellular localization. An OsDof-13 over-expression vector was constructed and transformed into a japonica rice variety Hejiang 19 by A grogacterium-mediated transformation. Southern blot assay indicated that OsDof-13 was successfully integrated into the Hejiang19 genome with 1-4 gene copies.