玉米BTB-TAZ蛋白与转录因子ZmBET10的互作分析

Analysis of Interaction between Transcription Factors ZmBET10 and BTB-TAZ Protein in Maize(Zea mays)

为明确玉米BTB-TAZ蛋白与转录因子ZmBET10的互作关系,本研究构建了玉米BTB-TAZ蛋白ZmBT4、Zm BT2a、ZmBT2b和转录因子ZmBET10(ZmBET10a,ZmBET10b)的酵母双杂交载体,利用酵母双杂交技术研究玉米BTB-TAZ蛋白与转录因子ZmBET10的互作关系。结果发现,共转化AD-ZmBT4、AD-ZmBT2a、AD-ZmBT2b分别与BD-ZmBET10a、BD-ZmBET10b组合的酵母在三缺培养基(SD/-Leu/-Trp/-His)和三缺添加3-AT的培养基(SD/-Leu/-Trp/-His/3-AT)上均能正常生长,而对照组均不能在三缺、三缺添加3-AT的培养基上正常生长,表明玉米BTB-TAZ蛋白ZmBT4、Zm BT2a、Zm BT2b均能与ZmBET10转录因子ZmBET10a、Zm BET10b在酵母细胞中直接互作。为明确BTB结构域在BTB-TAZ蛋白与转录因子ZmBET10互作中的功能,本研究构建了ZmBT4、ZmBT2b蛋白BTB结构域的酵母双杂交载体,进一步利用酵母双杂交技术研究BTB结构域与转录因子ZmBET10的互作,结果发现ZmBT4、Zm BT2b蛋白的BTB结构域ZmBT4-BTB、Zm BT2b-BTB均能与ZmBET10互作,表明BTB结构域在BTB-TAZ蛋白与转录因子ZmBET10互作过程中具有重要作用。研究结果为阐明BTB-TAZ蛋白在玉米抗病中的功能与调控机制奠定了基础。

In order to clarify the interaction between BTB-TAZ protein and transcription factor Zm BET10,the yeast two-hybrid vectors of BTB-TAZ protein ZmBT4,Zm BT2a,ZmBT2b and transcription factors ZmBET10(ZmBET10a and ZmBET10b)were constructed in this study.Yeast two-hybrid technology was used to study the interaction between BTB-TAZ protein and ZmBET10.The results showed that the yeast colonies that co-transformed with AD-ZmBT4,AD-ZmBT2a,AD-ZmBT2b combined with BD-ZmBET10a,BD-ZmBET10b could grow normally on three deficient medium(SD/-Leu/-Trp/-His)and three deficiency plus 3-AT(SD/-Leu/-Trp/-His/3-AT),while the control combinations could not grow normally,indicating that Zm BT4,Zm BT2a and Zm BT2b can directly interact with ZmBET10 in yeast cells.In order to clarify the function of BTB domain in the interaction between BTB-TAZ protein and transcription factor ZmBET10,yeast two hybrid vectors of BTB domain of Zm BT4,and ZmBT2b proteins were constructed.The interaction between BTB domain and transcription factor Zm BET10was further studied by yeast two hybrid technique.The results showed that BTB domain of ZmBT4 and ZmBT2b proteins could interact with ZmBET10,indicating that BTB domain plays an important role in the interaction between BTB-TAZ protein and transcription factor ZmBET10.These results laid a foundation for further study on the function and mechanism of BTB-TAZ protein in maize disease resistance.