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为了筛选化学杂交剂苯哒嗪的靶标蛋白基因,以SQ-1(15%苯哒嗪乳油)处理的小麦品种西农1376为材料,以噬菌体λTripl Ex2为载体,利用SMART技术构建小麦单核期花药c DNA表达文库;以生物素为报告基团,合成苯哒嗪小分子探针,并利用该探针在c DNA表达文库中进行苯哒嗪靶标蛋白基因筛选。结果成功构建了一种新型有效的药物印迹筛选方法,获得了一条能够与苯哒嗪结合的蛋白基因c DNA序列,其编码蛋白属于DUF3456家族。研究结果为进一步明确SQ-1(有效成分苯哒嗪)的作用机理提供了依据。
In order to screen the target protein gene of benzodiazepine, SQ-1 (15% benzodiazepine EC) -treated wheat cultivar Xinong 1376 was used as material, phage λTripl Ex2 was used as a carrier, and the wheat mononuclear stage Anther c DNA expression library; with biotin as a reporter group, to synthesize tribenzopyrazine small molecule probe, and using the probe in c DNA expression library phenylazine target protein gene screening. Results A novel and effective method of Western blotting was successfully constructed and a c DNA sequence of protein gene binding to tribenzyl was obtained. The encoded protein belonged to DUF3456 family. The results provide a basis for further clarifying the mechanism of action of SQ-1 (the active ingredient).