铁相关蛋白Dps对迟缓爱德华氏菌毒力影响的研究

发布时间：2018-01-07 08:37

  本文关键词：铁相关蛋白Dps对迟缓爱德华氏菌毒力影响的研究　出处：《中国科学院研究生院（海洋研究所）》2011年硕士论文　论文类型：学位论文

  更多相关文章： 迟缓爱德华氏菌 铁蛋白 Dps(DNA-binding proteins from starved cells)

【摘要】：Dps(DNA-binding proteins from starved cells)属于铁蛋白超家族,是原核生物中特有的一类能结合铁离子、抗氧化损伤和保护DNA的重要功能蛋白。本研究在迟缓爱德华氏菌株TX01中克隆得到了Dps家族的两个基因dps1和dps2。将dps1和dps2基因在大肠杆菌中重组表达后,获得的可溶性蛋白rDps1和rDps2具有DNA结合功能和铁氧化酶活性。为了检测dps1和dps2的功能作用,我们构建了dps1和dps2的单基因缺失突变株TXDps1和TXDps2以及突变菌株TXDps2的dps1基因抑制表达菌株TXDps2RI。与野生型菌株TX01相比,TXDps1和TXDps2的生长没有显著性差异,但TXDps2RI的逆境生长能力降低;TXDps1、TXDps2及TXDps2RI的抗氧化损伤、抗紫外线辐射能力皆明显减弱,但TXDps2RI的减弱程度较TXDps1和TXDps2更显著。毒力分析表明,与野生型菌株TX01以及TXDps1和TXDps2相比,TXDps2RI在牙鲆PBL中的生存复制能力显著降低,对牙鲆组织的感染能力大幅下降。综上所述,本研究证明了Dps1和Dps2具有保护DNA和抗氧化、抗辐射功能,并与迟缓爱德华氏菌致病力密切相关。
[Abstract]:Dps(DNA-binding proteins from starved cells belong to the superfamily of ferritin. It is a special kind of binding iron ions in prokaryotes. In this study, two genes of Dps family, dps1 and dps2, were cloned from TX01 of Edwards tardy strain. Dps1 and dps were cloned. 2 gene was expressed in E. coli. The soluble protein rDps1 and rDps2 had DNA binding function and iron oxidase activity. In order to detect the function of dps1 and dps2. The single gene deletion mutants TXDps1 and TXDps2 of dps1 and dps2 and the dps1 gene inhibitory expression strain TXDps2RIof the mutant TXDps2 were constructed. Compared with wild-type strain TX01. There was no significant difference between the growth of TXDps1 and TXDps2, but the growth ability of TXDps2RI was decreased under stress. TXDPS1, TXDPS2 and TXDps2RI were significantly reduced in anti-oxidative damage and anti-ultraviolet radiation ability. But the decrease of TXDps2RI was more significant than that of TXDps1 and TXDps2. Virulence analysis showed that compared with wild-type strains TX01, TXDps1 and TXDps2. The survival and replication ability of TXDps2RI in PBL was significantly lower than that in flounder tissues. This study demonstrated that Dps1 and Dps2 have protective effects on DNA, antioxidant and radiation resistance, and are closely related to the pathogenicity of Edwardsiella tardy.
【学位授予单位】：中国科学院研究生院（海洋研究所）
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R378

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