摘要
DREB(dehydration responsive element binding)转录因子在小麦非生物胁迫中起着非常重要的作用,但由于目前缺乏可识别小麦内源性DREB蛋白的抗体,导致其在蛋白水平上的研究进展非常缓慢。本研究通过分析DREB4A、4B和4C三种蛋白序列,将DREB4A在大肠杆菌中进行表达,并利用纯化后的蛋白作为抗原免疫兔子,在国内外首次获得小麦DREB4的多克隆抗体。Western blot结果证明,该抗体可特异性识别小麦内源性DREB4蛋白。该抗体介导的免疫组织化学结果显示,DREB4蛋白定位于细胞核内。本研究为深入研究植物DREB信号通路提供了有力的检测工具。
DREB(dehydration-responsive element-binding)transcription factor plays a very important role in wheat abiotic stress.However,due to the lack of antibodies that can recognize wheat endogenous DREB protein,its research progress at protein level is very slow.In this study,by analyzing three protein sequences of DREB4A,4B and 4C,the DREB4A was selected to express in Escherichia coli,and the purified protein was used as an antigen to immunize rabbits,then the polyclonal antibody of wheat DREB4 was obtained for the first time at home and abroad.The Western blot results showed that the antibody could specifically recognize wheat endogenous DREB4 protein.The antibody-mediated immunohistochemical results showed that DREB4 protein was localized in the nucleus.This study could provide a powerful detection tool for in-depth research of plant DREB signaling pathway.
作者
李永波
鲁琳
方会见
崔德周
孟福燕
黄琛
隋新霞
樊庆琦
楚秀生
Li Yongbo;Lu Lin;Fang Huijian;Cui Dezhou;Meng Fuyan;Huang Chen;Sui Xinxia;Fan Qingqi;Chu Xiusheng(Crop Research Institute,Shandong Academy of Agricultural Sciences/Key Laboratory of Wheat Biology and Genetics and Breeding in Northern Huang-Huai River Plain,Ministry of Agriculture and Rural Affairs/Shandong Technology Innovation Center of Wheat/Jinan Key Laboratory of Wheat Genetic Improvement,Jinan 250100,China;Shandong Luyan Seed Co.,Ltd.,Jinan 250100,China;Yuncheng Country Seed Company,Yuncheng 274700,China;College of Life Sciences,Yantai University,Yantai 264000,China)
出处
《山东农业科学》
北大核心
2023年第3期9-14,共6页
Shandong Agricultural Sciences
基金
山东省农业良种工程项目(2019LZGC01702)
山东省农业良种工程项目(2021LZGC013)
山东省自然科学基金青年项目(ZR2020QC114)
国家自然科学基金青年项目(32001542)
小麦玉米国家工程实验室开放课题(2018LYZWS06)。
