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为了阐明转录激活因子FleQxoo对水稻白叶枯病菌(Xanthomonas oryzaepv.oryzae,Xoo)鞭毛基体基因fliExoo的转录调控机理,本研究用PCR方法从Xoo野生型菌株PXO99A中克隆了fleQxoo基因,构建了原核表达载体pET21-fleQ-xoo,在大肠杆菌BL21(DE3)pLysS中进行了诱导表达,通过Ni-NTA亲和层析纯化得到FleQxoo蛋白;EMSA检测到FleQ-xoo与鞭毛基体基因启动子fliExoo-p片段的特异结合作用。这些结果为阐明FleQxoo直接调控鞭毛基体基因fliExoo的转录提供了分子证据。
In order to elucidate the transcriptional regulatory mechanism of the flagellar gene fliExoo of Xanthomonas oryzaepv. Oryzae (Xoo), a FleQxoo gene was cloned from Xoo wild type strain PXO99A by PCR to construct a prokaryotic expression vector The vector pET21-fleQ-xoo was induced in E. coli BL21 (DE3) pLysS and the purified FleQxoo protein was purified by Ni-NTA affinity chromatography. EMSA detected the fliExoo-p fragment of FleQ-xoo and flagella matrix gene promoter The specific binding. These results provide molecular evidence for clarifying that FleQxoo directly regulates the transcription of the flagella matrix gene fliExoo.