CAJ | 학술논문

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(Xanthomonas oryzae pv.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(Xoryzae pv.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.
근거황단포균hisF기인적동원성설계간병인물,채용PCR방법종수도백협고병균(Xanthomonas oryzae pv.oryzae,Xoo)중극륭료hisF동원기인,명명위hisFXoo.NCBI망참단백질보수결구역수색표명,HisFXoo속우조안산생물합성단백가족적일원,구유여기타HisF단백상사적결구.서렬비교현시,해기인재황단포균중시상대보수적.동원성수색비교현시,여수도조반병균(Xoryzae pv.oryzicola,Xooc)적hisF(등록호:DQ372107)동원성고체98%.통과동원중조적방법,구건료hisFXoo적삽입돌변체,재NA+Amp평판상사선후,병차경과PCR급Southern잡교험증삽입정학.재MMX기본배양기중생장측정발현,hisF돌변체생장능력명현하강,증명hisF기인여Xoo생장대사상관.단시,해돌변체잉구유치병성,병차독력여야생형균주몰유명현차이.
A hisF homologue, designated hisFXoo, was cloned from the genomic DNA of Xanthomonas oryzae pv. oryzae (Xoo) with degenerated primers by PCR. NCBI blast search indicated that HisEXoo had a similar structure to that of other HisF proteins, and had conserved domain of histidine biosynthesis protein. Nucleotide sequence comparison showed that hisFXoo was conserved in the Xanthomonas genus. Homology search revealed that the hisFXooc was 98% similarities to hisF ( Accession number: DQ372107) of X. oryzae pv. oryzicola (Xooc). A hisFXoo disruption mutant was successfully generated by a single cross-over e-vent, selected by Amp resistance on NA medium, and confirmed by PCR and Southern blot. The growth of the mutants was reduced compared to wild type strain when grown in MMX medium. The results suggest that hisFXoo is involved metabolism of Xoo. However, the mutant still had strong pathogenicity, and its virulence was not obviously different from that of wild type strain. Nevertheless, how hisF to regulate metabolism of Xoo, transcription and expression of genes involved in metabolism still need to be further studied.