利用药物印迹法筛选化学杂交剂苯哒嗪靶标蛋白基因

Screening target-protein gene of chemical hybridizing agent clofencet by drug-western method

摘要: 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λTriplEx2为载体,利用SMART技术构建小麦单核期花药cDNA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cDNA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cDNA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。

Abstract: In order to select the target gene of chemical hybridizing agent clofencet, the cDNA expression library of uninucleate-stage anther from wheat varieties Xinong 1376 treated with SQ-1 (15% clofencet EC) was constructed by SMART technology using the carrier phage λ TriplEx2. The clofencet target genes were screened by a synthetic small molecule probes with a reporter group biotin. A novel method for effective drug screening blots was set up, and a target gene of clofencet was obtained, which encoded a protein belongs to the DUF3456 family. The results provided basis for further study on the mechanisms of SQ-1 (the active ingredient of clofencet).