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为研究Gm SPX3在大豆低磷胁迫响应中的作用机制,寻找与Gm SPX3发生相互作用的蛋白质,利用Gateway技术构建了低磷大豆根系酵母杂交c DNA文库,并使用p GBKT7-Gm SPX3融合蛋白为诱饵,采用酵母双杂交方法筛选酵母杂交c DNA文库,经过显色反应验证后获得21个阳性克隆,通过生物信息学分析,选择2个转录因子编码基因Gm UNE12和Gm Pos F21作为候选基因进行克隆,并分别构建AD载体p GADT7-Gm UNE12和p GADT7-Gm Pos F21,与诱饵p GBKT7-Gm SPX3进行一对一互作验证,结果证明Gm UNE12和Gm Pos F21蛋白均与Gm SPX3诱饵蛋白发生了相互作用。
In order to study the mechanism of Gm-SPX3 in response to low-P stress in soybean, we searched for the protein interacting with Gm-SPX3 and constructed the low-phosphorus Soybean hybrid yeast c DNA library by Gateway technology. Using p GBKT7-Gm SPX3 fusion protein Bait and yeast two-hybrid method were used to screen yeast hybrid c DNA library. After verification by chromogenic reaction, 21 positive clones were obtained. Two biomarkers including Gm UNE12 and Gm Pos F21 were selected as candidate genes by bioinformatics analysis. , And the AD vectors pGADT7-Gm UNE12 and pGADT7-GmPOS F21 were constructed and verified by one-to-one interaction with bait pGBKT7-Gm SPX3, respectively. The results showed that both Gm UNE12 and Gm Pos F21 proteins were associated with Gm SPX3 bait protein Interaction.