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OALib Journal期刊
ISSN: 2333-9721
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播娘蒿转录因子dscbf与crt/dre元件的相互作用
DOI: 10.3724/SP.J.1145.2010.00609, PP. 609-612
Keywords: 播娘蒿,cbf,crt/dre,emsa,银染
Abstract:
cbf（c-repeatbindingfactor）是调控植物冷驯化相关基因表达的一种调控转录激活因子，cbf蛋白ap2结合domain可与cor基因启动子cbt/dre元件特异性地结合，启动耐寒基因cor的表达.为研究播娘蒿dscbf的功能，构建pet32a-dscbf原核表达载体，通过冻融法转化入大肠杆菌bl21（de3）中，进行原核表达，并通过镍亲和层析纯化cbf蛋白.利用emsa（electrophoreticmobilityshiftassay）分析dscbf蛋白与播娘蒿dscor基因的启动子上crt/dre元件的相互作用.emsa结果显示，dscbf蛋白与含有ccgac核心序列及tata-box和at-ta回文结构的50bp探针结合有滞后现象，表明与dscor启动子上的crt/dre元件有特异性结合；而与仅有ccgac的crt/dre核心序列的40bp探针结合则无滞后现象，表明dscbf与蛋白结合的识别可能与tata-box和at-ta回文结构有关.图6参19
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