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AAA ATPases(ATPase associated with diverse cellular activities)是一种与细胞活性相关的酶,广泛存在于所有生物中,同时具有蛋白酶和分子伴侣活性。为了研究AAA ATPases相关基因缺失对西瓜噬酸菌(Acidovorax citrulli)的影响,本研究通过同源重组双交换的方法构建西瓜噬酸菌moxR基因以及ruv B基因全缺失突变菌株及互补菌株,测定了突变菌株的致病性、烟草过敏反应、运动性、生长能力、生物膜形成等;并利用实时荧光定量PCR方法测定了AAA ATPases与Ⅲ型分泌系统相关基因(hrc N和hrp E)、毒性蛋白相关基因(trb C和vir B)、鞭毛相关基因(fliR和fli M)、菌毛相关基因(pin C-1和pin C-2)以及AAA ATPases亚家族Ruv B相关基因(ruv A和ruv C)的关系。结果表明,moxR基因以及ruv B基因全缺失突变菌株的致病力降低、运动性减弱、生长能力下降、生物膜形成能力增强、烟草过敏性反应减弱、突变菌株中重要的致病基因hrc N和hrp E表达量显著下调,表明AAA ATPases相关基因moxR、ruv B在西瓜噬酸菌的致病力上发挥重要作用。
AAA ATPases (ATPase associated with diverse cellular activities) is a cell-activity-related enzyme found in all living organisms and has both protease and chaperone activities. In order to study the influence of AA ATPase-related gene deletion on Acidovorax citrulli, moxR gene of watermelon phage and full deletion mutant of ruv B gene and its complementary strains were constructed by homologous recombination double exchange. Pathogenicity of the mutant strain, allergic reaction to tobacco, motor activity, growth ability, biofilm formation, etc. Real-time fluorescence quantitative PCR was used to determine the association between AAA ATPases and the type Ⅲ secretion system related genes (hrc N and hrp E) Related genes (trb C and vir B), flagella-related genes (fliR and fli M), pilus-related genes (pin C-1 and pin C-2) and AAA ATPases subfamilies Ruv B related genes (ruv A and ruv C )Relationship. The results showed that the moxR gene and ruv B gene deletion mutant had lower pathogenicity, weakened motility, decreased growth ability, enhanced biofilm formation ability and weakened tobacco allergic reaction. The genes of hrc N and hrp E was significantly downregulated, indicating that AAA ATPases related genes moxR, ruv B in the pathogenicity of watermelon bacteria play an important role.