利用药物印迹法筛选化学杂交剂苯哒嗪靶标蛋白基因

赵新亮; 张改生; 宋瑜龙; 李志宽; 朱启迪; 赵卓军; 马守才; 王军卫; 牛娜

doi:10.3969/j.issn.1008-7303.2014.06.13

利用药物印迹法筛选化学杂交剂苯哒嗪靶标蛋白基因

doi: 10.3969/j.issn.1008-7303.2014.06.13

1.
西北农林科技大学/国家杨凌农业生物技术育种中心/国家小麦改良中心杨凌分中心/小麦育种教育部工程研究中心/陕西省作物杂种优势研究与利用重点实验室, 陕西杨凌 712100
2.
河南科技学院生命科技学院, 河南新乡 453003

基金项目:

国家高技术研究发展计划(“863”计划)重大专项(2011AA10A106);国家自然科学基金项目(31171611, 31371697);陕西省 “13115”科技创新工程与科技统筹重大科技专项(2010ZDKG-68,2014KTZB02-01-02).

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出版历程
- 收稿日期: 2014-07-31
- 修回日期: 2014-10-10
- 刊出日期: 2014-12-12

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

1.
Northwest Agriculture and Forestry University/National Breeding Center of Agricultural Biotechnology in Yangling/Yangling Branch of State Wheat Improvement Center/Wheat Breeding Engineering Research Center, Ministry of Education/Key Laboratory of Crop Heterosis of Shaanxi Province, Yangling 712100, Shaanxi Province, China
2.
Department of Life Science and Technology, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China

摘要

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。
- 小麦 /
- 化学杂交剂 /
- SQ-1 /
- 苯哒嗪 /
- 药物印迹 /
- cDNA表达文库 /
- 靶标蛋白基因
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).
- wheat /
- chemical hybridizing agent /
- SQ-1 /
- clofencet /
- drug-western method /
- cDNA expression library /
- target-protein gene

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参考文献(24)

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