油菜转录因子BnZFP基因的分离及其功能初步分析

Cloning and Preliminary Characterization of the BnZFP Gene from Brassica napus

为克隆油菜ZFP类转录因子基因,初步阐明其功能。采用RT-PCR的方法克隆基因和体内检测;花沾法转化拟南芥,Northernblotting检测转基因后的表达。从甘蓝型油菜中分离了一个新的ZFP类转录因子,它编码377个氨基酸,与拟南芥AtZFP基因(At2g29660)的碱基序列相似性最高,为76.2%,命名为BnZFP。该基因在父本、母本和F1代不同发育时期中均能较稳定的表达;该基因转化拟南芥,对获得的转BnZFP基因拟南芥进行了NorthernBlotting分析,表明该基因在拟南芥中得到过表达;与野生型拟南芥相比较,过表达BnZFP的转基因拟南芥在萌发、营养生长及生殖生长过程中没有表现出明显的表型变化,该基因的功能有待进一步研究。

To clone a new transcription factor from Brassica napus, and study its function, methods including semi-quantitative RT-PCR, transformation, Northern-blotting were used. BnZFP was isolated from Brassica.napus, which constituted 377 amino acids. BnZFP had the highest similarity with AtZFP at nucleotide and amino acids level. Semi-quantitative RT-PCR showed BnZFP was steadily expressed throughout the different developmental stages between F1 and parental lines. BnZFP was constructed to pBI121 vector and this expression cassette was introduced into wild Arabidopsis thaliana. The transgenic plants were analysed by Northern blotting. Compared with wild Arabidopsis thaliana, over-expression of BnZFP in plants didn＇t show any visible changes in seed germination and post-germination growth, further study should be carried out to study the gene＇s function.