水稻白叶枯病菌gacA基因的克隆、测序及敲除研究  

Cloning,sequencing and knock-out of gacA gene from Xanthomonas oryzae pv.oryzae

作　　者：杨万风[1] 刘红霞[1] 娄旸[1] 胡白石[1] 许志刚[1] 刘凤权[1] 

机构地区：[1]南京农业大学农业部病虫监测与治理重点开放实验室,江苏南京210095

出　　处：《南京农业大学学报》2006年第3期33-38,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(30070497);国家高技术研究发展计划(2002AA245041)

摘　　要：根据黄单胞菌gacA基因的同源性设计简并引物,采用PCR方法从水稻白叶枯病菌(Xanthom onas oryzaepv.oryzae,Xoo)中克隆了gacA同源基因,命名为gacAXoo。序列比较显示,该基因在黄单胞菌中是相对保守的。蛋白质保守结构域搜索表明,GacAXoo属于LuxR家庭的一员,具有与其他GacA蛋白相似的结构。通过同源重组的方法,构建了gacAXoo的插入突变体,为下一步研究gacAXoo的功能奠定了良好的基础。A gacA homologue, designated gacAxoo, was cloned from the genomic DNA of Xanthornonas oryzae pv. oryzae （Xoo）, a bacterium that causes bacterial blight of rice, with degenerated primers by polymerase amplification reaction （PCR）. NCBI blast search indicated that GacAxoo had a similar structure to that of other GacA proteins, and had conserved domain of LuxR-like proteins. Sequence comparison showed that gacAxoo was conserved in the Xanthomonas genus. A gacAxoo disruption mutant was successfully generated by a single cross-over event, selected by Amp resistance on NA medium, and confirmed by PCR and Southern blot. This mutant is a useful tool for further study on the function of gacAxoo in X. oryzae pv. oryzae.

关 键 词：水稻 白叶枯病菌 gacA基因 基因敲除 

分 类 号：S435.111[农业科学—农业昆虫与害虫防治]