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水稻白叶枯病菌hisF基因的克隆及功能初析

DOI: 10.7685/j.issn.1000-2030.2009.04.014, PP. 72-78

Keywords: 水稻白叶枯病菌,hisF基因,突变体,生长代谢

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Abstract:

根据黄单胞菌hisF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(Xanthomonasoryzaepv.oryzae,Xoo)中克隆了hisF同源基因,命名为hisFXoo。NCBI网站蛋白质保守结构域搜索表明,HisFXoo属于组氨酸生物合成蛋白家族的一员,具有与其他HisF蛋白相似的结构。序列比较显示,该基因在黄单胞菌中是相对保守的。同源性搜索比较显示,与水稻条斑病菌(X.oryzaepv.oryzicola,Xooc)的hisF(登录号DQ372107)同源性高达98%。通过同源重组的方法,构建了hisFXoo的插入突变体,在NA+Amp平板上筛选后,并且经过PCR及Southern杂交验证插入正确。在MMX基本培养基中生长测定发现,hisF突变体生长能力明显下降,证明hisF基因与Xoo生长代谢相关。但是,该突变体仍具有致病性,并且毒力与野生型菌株没有明显差异。

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