转录激活因子FleQxoo原核表达及其与鞭毛基体基因fliExoo启动子的结合  

A transcriptional activator FleQxoo binds to promoter of fliExoo,a flagellar base body gene in Xanthomonas oryzae pv. oryzae

作　　者：傅本重[1] 吴茂森[1] 陈华民[1] 何晨阳[1] 

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193

出　　处：《植物病理学报》2009年第5期476-481,共6页Acta Phytopathologica Sinica

基　　金：中央财政国家重点实验室自主研究课题专项(SKL2007SR06)

摘　　要：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。To elucidate the transcriptional regulation mechanism of flagellar genes by transcriptional activator FleQxoo in Xanthomonas oryzae pv. oryzae （Xoo）, the fleQxoo gene was cloned from the wild-type strain PXO99A of Xoo by PCR and the expression plasmid pET21-fleQxoo was constructed. The FleQxoo protein was expressed in Escherichia coli strain BL21 （DE3）pLysS with IPTG induction and purified through Ni-NTA affinity chromatography. The purified FleQxoo could be directly bound to the DNA fragments of fliExoo promoter in vitro by EMSA （Electrophoretic Mobility Shift Assays）. The results provided the evidence that FleQxoo might regulate the flagellar gene expression by direct binding to the promoters in Xoo.

关 键 词：水稻白叶枯病菌 FleQxoo 原核表达 fliExoo EMSA 

分 类 号：S435.111.47[农业科学—农业昆虫与害虫防治]