流产布鲁菌毒力相关基因鉴定及LsrB蛋白功能研究

发布时间：2018-06-22 00:13

本文选题：流产布鲁菌 + 毒力相关基因　；参考：《中国农业科学院》2016年博士后论文

【摘要】：本研究在前期PCR介导的信号标签随机突变库基础上,以感染小鼠为模型,筛选鉴定流产布鲁菌毒力相关基因。为进一步研究布鲁菌毒力相关基因的功能,构建了一系列用于布鲁菌克隆和表达外源基因的质粒。之后,对布鲁菌一个LysR家族调控子的功能进行详细阐明,为布鲁菌致病机制研究奠定基础。本文的主要结论是：1.共获得56株流产布鲁菌减毒突变株,鉴定53个毒力相关基因,其中,39个是本研究中新鉴定的基因。2.注释53个毒力相关基因,功能分类探讨相关基因可能与布鲁菌毒力之间的关系。3.构建一系列pTM质粒用于在布鲁菌中克隆外源基因,包括pTM1-Cm、pTM2-Amp和pTM3-Km。4.构建pGH-6x His和pZT-6x His质粒,为布鲁菌表达和诱导性表达外源基因提供条件。5.鉴定一个与布鲁菌毒力相关的LysR家族转录调控子LsrB蛋白,参与调控布鲁菌胞内存活和建立慢性感染。6.发现LsrB调控布鲁菌抵抗氧化应激、氮化应激、自然血清以及调控布鲁菌逃逸宿主免疫监控。7.揭示LsrB在根瘤菌目细菌中具有保守的共同调控机制。
[Abstract]:On the basis of the early PCR mediated random mutation library, this study screened the Virulence Related Genes of Brucella abortus using the infected mice as the model. In order to further study the function of Brucella virulence related genes, a series of plasmid for Brucella cloning and expression of foreign genes was constructed. Then, a LysR family of Brucella was used. The function of the regulator was clarified in detail to lay the foundation for the study of the pathogenic mechanism of Brucella. The main conclusions of this paper were: 1. a total of 56 strains of Brucella abortus were obtained and 53 virulence related genes were identified. Among them, 39 were.2. annotated and 53 Virulence Related Genes of the newly identified gene in this study, and the functional classification was used to explore the possible genes. The relationship with Brucella virulence.3. constructs a series of pTM plasmids to clone foreign genes in Brucella, including pTM1-Cm, pTM2-Amp and pTM3-Km.4. to construct pGH-6x His and pZT-6x His plasmids, providing conditions for the expression and inducible expression of foreign genes for Brucella, and.5. to identify a LysR family transcriptional regulator associated with Brucella virulence SrB protein, participating in the regulation of Brucella cell memory and the establishment of chronic infection.6. found that LsrB regulated Brucella to resist oxidative stress, nitridation stress, natural serum and control of the immune monitoring of Brucella escaping host immune monitoring.7. revealed that LsrB has a conservative control mechanism in Rhizobium bacteria.
【学位授予单位】：中国农业科学院
【学位级别】：博士后
【学位授予年份】：2016
【分类号】：Q93

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