發(fā)布時間：2021-07-24 09:51

　　PhoP/PhoQ系統(tǒng)是鼠傷寒沙門氏菌毒力調(diào)控中最關(guān)鍵的二元調(diào)控系統(tǒng)之一。已知的可以激活PhoP/PhoQ系統(tǒng)的信號包括低濃度的鎂離子,酸性pH,以及陽離子抗菌多肽（cAMP） （1,2）。PhoP/PhoQ系統(tǒng)被激活后,可以調(diào)節(jié)100多個基因的表達（3）,同時PhoP/PhoQ系統(tǒng)對一些其它的二元調(diào)控系統(tǒng),如SsrA/SsrB（4）, PmrA/PmrB（5）和RstA/RstB（6）也有調(diào)控作用。有研究表明,沙門氏菌中RstB （RstA/RstB系統(tǒng)中的組氨酸激酶）可以與PhoQ相互作用從而激活PhoP/PhoQ（7）。另外,RstA也可以在低濃度鎂離子或酸性pH條件下被PhoP/PhoQ系統(tǒng)激活（8）。這些研究說明在關(guān)鍵調(diào)控系統(tǒng)PhoP/PhoQ和了解較少的RstA/RstB系統(tǒng)之間可能存在著關(guān)聯(lián)和互作。通過系統(tǒng)生物學(xué)方法研究PhoP/PhoQ系統(tǒng)的工作很多,而對RstA/RstB系統(tǒng)調(diào)控網(wǎng)絡(luò)的系統(tǒng)性研究目前還沒有報道。為了更好地解析RstA/RstB系統(tǒng)以及RstA/RstB 與 PhoP/PhoQ之間的關(guān)系,我們采用了一個已知的PhoP/PhoQ和RstA/RstB的激活條... 

【文章來源】：武漢大學(xué)湖北省 211工程院校 985工程院校 教育部直屬院校

【文章頁數(shù)】：153 頁

【學(xué)位級別】：博士

【文章目錄】：
Abstract
中文摘要
Chapter 1:Introduction
    1.1 Mass spectrometry-based proteomics
        1.1.1 The concept of proteomics
        1.1.2 Mass spectrometry basics
        1.1.3 Mass spectrometry-based proteomics technique
        1.1.4 Quantitative proteomics
        1.1.5 Data analysis and bioinformatics tool
    1.2 Salmonella pathogenesis
        1.2.1 Salmonella overview
        1.2.2 Two-component system
        1.2.3 The application of proteomics in Salmonella pathogenesis
    1.3 Research hypothesis and aims
Chapter 2:Proteomics Analysis of RstA/RstB System in Salmonella Typhimurium
    2.1 Introduction
    2.2 Materials and methods
        2.2.1 Bacterial strains,medium and growth conditions
        2.2.2 Protein sample preparation
        2.2.3 Two-dimensional electrophoresis methods
        2.2.4 Protein digestion
        2.2.5 Peptide stable isotope dimethyl labeling
        2.2.6 Peptide separation by strong cation exchange(SCX)chromatography
        2.2.7 Mass spectrometry analysis and data processing
        2.2.8 Bioinformatics analysis
        2.2.9 Quantitative real-time PCR(qPCR)analysis
        2.2.10 Phenotype analysis
    2.3 Results and discussion
        2.3.1 Experimental strategies and proteomics workflow
        2.3.2 Overview of the proteomics data
        2.3.3 Metabolic pathways and function groups regulated by RstA/RstB
        2.3.4 RstA/RstB TCS regulation of pyrimidine metabolism
        2.3.5 RstA/RstB TCS regulation of iron acquisition
        2.3.6 RstA/RstB regulation of surface motility, chemotaxis and invasion
    2.4 Conclusion
Chapter 3:Proteomics Analysis of PhoP/PhoQ System in SalmonellaTyphimurium
    3.1. Introduction
    3.2. Materials and methods
        3.2.1. Bacterial strains, medium and growth conditions
        3.2.2. Proteins sample preparation
        3.2.3 Biofilm phenotype analysis
    3.3. Results and discussion
        3.3.1 Experimental strategies and proteomics workflow
        3.3.2 Overview of the proteomics data
        3.3.3 Metabolic pathways and function groups regulated by PhoP/PhoQ system
        3.3.4 Network analysis of proteins regulated by PhoP/PhoQ system
        3.3.5 PhoP/PhoQ regulates several two-component systems
        3.3.6 PhoP/PhoQ regulation of ribosome proteins
        3.3.7 PhoP/PhoQ regulates ABC transport proteins
        3.3.8 phoPQ mutant effected to biofilm formation
    3.4. Conclusion
Chapter 4:Comparative Proteomics Analysis of Salmonella Typhimurium underthe Regulation of RstA/RstB and PhoP/PhoQ Systems
    4.1 Introduction
    4.2 Materials and Methods
    4.3 Results and discussion
        4.3.1 Comparative proteomics data overview of the two mutant strains
        4.3.2 Comparison of metabolic pathways and functional groups significantly regulated by RstA/RstB and PhoP/PhoQ TCSs
        4.3.3 Comparison of common proteins regulated by RstA/RstB and PhoP/PhoQ TCSs
    4.4 Conclusion
Chapter 5:Summary and Future Perspective
References
Supplemental S1.80 metabolic pathways of △rstB and △phoPQ strains
Supplemental S2.Changed protein of △rstB strain
Supplemental S3.Changed protein of △phoPQ strain
Research Publications
Acknowledgements



本文編號：3300424