大豆GmNF-YA13蛋白是一个核转录因子Y(NF-Y),在干旱和高盐响应过程中均发挥重要作用.为研究其抗旱和耐盐的作用机理,寻找GmNF-YA13的互作蛋白,构建pGBKT7-GmNF-YA13诱饵载体,采用酵母双杂交筛选大豆酵母文库,并进行X-α-gal染色验证.结果显示:酵母双杂交获得85个阳性克隆,测序分析后得到36个候选的互作蛋白.功能预测显示互作蛋白主要参与生长发育、胁迫响应、能量代谢、转录调控和信号转导等生物过程.选择GmUVR8、GmCML41、GmFbox13 和 GmFBA 与诱饵 pGBKT7-GmNF-YA13 进行一对一验证,只有 GmFBA 能与 GmNF-YA13发生相互作用,预示GmNF-YA13功能的发挥需要GmFBA的参与.该结果可为NF-YA抗逆分子网络的研究提供基础.
Soybean(Glycine max)GmNF-YA13 protein is a nuclear transcription factor Y(NF-Y)that plays a crucial role in drought and high salinity responses.To investigate its mechanisms of drought resistance and salt tolerance,and identify the interacting proteins of GmNF-YA13,a pGBKT7-GmNF-YA13 bait vector was constructed.The soybean yeast library was screened using yeast two-hybrid system,and verified by staining with X-α-gal.The result showed that:85 positive clones were obtained after sequencing analysis,36 candidate interacting proteins were identified.Functional prediction showed that these interacting proteins are mainly involved in growth and development,stress responses,energy metabolism,transcription regulation,and signal transduction processes.GmUVR8,GmCMI41,GmFbox13,and GmFBA were selected for one-on-one validation with the bait vector pGBKTI7-GmNF-YA13.The results revealed that only GmFBA could interact with GmNF-YA13,indicating that the function of GmNF-YA13 requires the involvement of GmFBA.These findings lay the foundation for further research on the NF-YA stress resistance molecular network.
刘灿;于月华;倪志勇
新疆农业大学生命科学学院,新疆乌鲁木齐 830052新疆农业大学农学院,新疆乌鲁木齐 830052
核转录因子;GmNF-YA13;酵母双杂交;互作蛋白
nuclear transcription factor;GmNF-YA13;yeast two-hybrid;interacting protein
《大豆科学》 2024 (001)
21-28 / 8
国家自然科学基金(32160446);新疆维吾尔自治区自然科学基金(2022D01E14,2021D01A93).
10.11861/j.issn.1000-9841.2024.01.0021
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